ISSN: 2763-5724 / Vol. 05 - n 05 - ano 2025

USE OF CRISPR BASE EDITING AND PRIME EDITING IN NEUROLOGICAL

DISEASES

Milena Gaion Malosso1

Eriana de Souza Batalha2

Ivan Monteiro dos Santos3

Ricardo dos Santos Faria4

Abstract: Next-generation gene editing technologies, particularly base editing and prime editing,

have emerged as highly promising approaches for the treatment of neurological diseases driven by

pathogenic mutations and genomic instability. Unlike conventional CRISPR-Cas9 methods that

introduce double-strand breaks, these systems enable precise nucleotide conversions or guided

sequence replacements, thereby reducing off-target events and increasing safety for central nervous

system (CNS) applications. Recent studies demonstrate their applicability in cellular and animal

models of conditions such as Alternating Hemiplegia of Childhood, Huntington’s disease, and

repeat-associated ataxias, showing functional restoration, decreased somatic repeat expansion, and

improvement of neurological phenotypes. However, the clinical translation of these tools still faces

substantial challenges, including limitations in CNS delivery, target-dependent efciency, and the

need for comprehensive biosafety evaluation. Viral and nonviral platforms—such as optimized

AAVs, lipid nanoparticles, and virus-like particles—are under active development to overcome these

barriers. Additional gaps remain regarding editing durability, immunogenicity, and scalability. This

article provides an integrated analysis of the principles, preclinical applications, technical limitations,

and future perspectives of base editing and prime editing in neurological diseases, emphasizing their

1 Doutorado em Biotecnologia

2 Graduando em Biotecnologia

3 Mestre em odontologia

4 Mestre em Saúde da Família

ISSN: 2763-5724 / Vol. 05 - n 05 - ano 2025

transformative potential and the necessity of rigorous, safety-driven research.

Keywords: Precision Editing. Molecular Medicine. Neurogenetics. Gene Therapy, Viral Vectors.

INTRODUCTION

The rapid advancement of genome editing technologies has profoundly transformed the eld

of biotechnology and biomedical sciences, offering increasingly accurate, efcient, and safe approaches

to DNA manipulation (Devinsky et al., 2025). Among these tools, the CRISPR base editing and

the Prime Editing emerge as state-of-the-art strategies capable of promoting specic modications

in the genome without generating double-strand breaks, signicantly reducing unwanted effects

associated with traditional methods. These techniques open up new possibilities for the investigation

and correction of pathogenic genetic variants, especially in tissues of high biological complexity, such

as the central nervous system (MacLaren et al., 2020).

The importance of the theme stands out in view of the signicant global burden of neurological

diseases, which affect millions of people and present, in many cases, complex etiology and limited

therapeutic response. Diseases such as Alzheimer’s, Parkinson’s, hereditary ataxias, genetic epilepsies,

and various neuropathies associated with point mutations pose signicant medical and socioeconomic

challenges (Feigin et al., 2020). In this context, base editing and prime editing methodologies offer

promising alternatives to correct pathological variants with greater precision, expanding the potential

for personalized treatments and early interventions. In addition, the applicability of these technologies

in cellular and animal models contributes to a deeper understanding of the molecular mechanisms

underlying neurological diseases (Murray, Harrison and Scholeeld, 2025).

The guiding question of this work is: How have base editing and prime editing technologies

been applied in the study and correction of mutations associated with neurological diseases, and

what are their advances, limitations, and therapeutic perspectives? The rationale lies in the growing

scientic and clinical interest in approaches that enable safe and highly specic genetic interventions

ISSN: 2763-5724 / Vol. 05 - n 05 - ano 2025

in neural tissue, where cell regeneration is limited and the effects of mutations can be particularly

devastating. Recent literature demonstrates a rapid increase in the number of studies exploring these

technologies in the neurological context, reinforcing the need for a comprehensive and critical review

on the topic (Paul, Collins, and Lee, 2022).

Therefore, the general objective of this article is to analyze the state of the art of the use

of CRISPR base editing and Prime Editing in neurological diseases, highlighting applications,

challenges and future perspectives. Specic objectives include: (i) to describe the molecular principles

and functional differences between base editing and prime editing; (ii) review experimental and

preclinical studies that use these technologies in models of neurological diseases; (iii) discuss the

main therapeutic advances reported, as well as technical, ethical, and safety limitations; and (iv) point

out gaps in the literature and possible directions for future research aimed at the clinical application

of these tools in neurological conditions (Chiba-Falek et al, 2025a).

METHODOLOGY

This study was conducted as a systematic review strictly following the guidelines established

by the PRISMA (Preferred Reporting Items for Systematic Reviews and Meta-Analyses) protocol,

ensuring transparency, reproducibility and methodological robustness at all stages of the investigation.

The elaboration of the initial protocol included a clear denition of the guiding question, the eligibility

criteria and the search strategies, ensuring that the process of selecting the studies was objective and

consistent with the scope of the research.

Search strategy

The research question was structured based on the PICO model adapted for biotechnology

reviews: What are the applications, advances, and limitations of CRISPR base editing and prime

ISSN: 2763-5724 / Vol. 05 - n 05 - ano 2025

editing technologies in neurological diseases? Thus, the following criteria were dened: Population

(P) – studies involving neurological diseases of genetic origin; Intervention (I) – use of CRISPR base

editing or prime editing; Comparison (C) – not applicable due to heterogeneity of studies; Outcome

(O) – genetic correction, phenotypic modulation, safety and efcacy assessment. Based on these

elements, inclusion and exclusion criteria were outlined.

Inclusion and exclusion criteria

The inclusion criteria involved: (i) original articles published between 2015 and 2025, a period

corresponding to the development and consolidation of base editing and prime editing technologies;

(ii) studies conducted in cell models, organoids, animal models or human samples; (iii) research that

directly applied one of the aforementioned CRISPR technologies to investigate or correct mutations

related to neurological diseases; and (iv) texts published in English or Portuguese and available in

full. The exclusion criteria were: (i) narrative reviews, editorials, letters, and opinion reports; (ii)

studies that used only conventional CRISPR/Cas9 methods without base editing or prime editing;

(iii) studies that addressed diseases unrelated to the nervous system; and (iv) studies without adequate

methodological description.

Systematic search

It was performed in the PubMed, Scopus and Web of Science databases, using combinations

of controlled descriptors (MeSH) and free keywords, such as: “CRISPR base editing”, “prime editing”,

“neurological diseases”, “neurogenetic disorders”, “genome editing therapy”. Boolean operators AND

and OR were employed to broaden and rene the results. All searches were carried out between

August and September 2025. The identied records were imported into the Rayyan software for

duplicate removal and initial screening.

ISSN: 2763-5724 / Vol. 05 - n 05 - ano 2025

Selection of studies

It took place in two stages: (i) screening by title and abstract, carried out by two independent

reviewers; and (ii) complete reading of the eligible texts. Divergences were resolved by consensus or by

a third reviewer. The complete process of identication, screening, eligibility and nal inclusion was

documented in a PRISMA owchart, ensuring the traceability of decisions. In the end, the included

studies had their data systematically extracted in a matrix containing information on the experimental

model, target mutation, type of editing, results obtained, limitations reported, and contributions to

the eld. The methodological quality assessment was conducted using criteria adapted from the NIH

Quality Assessment Tool, allowing the studies to be classied for risk of bias

LITERATURE REVIEW

Technologies, principles and evolution (base editing vs. prime editing).

Base editing (BE) and prime editing (PE) have emerged as genome editing platforms

designed to increase the accuracy of modications without relying on double-strand breaks (DSBs)

and repair by homologous recombination with a donor template. The rst functional BE described

was the Cytosine Base Editor (CBE), which combines a cytosine-deaminase to the knocked out Cas9

variant (nickase or dCas9) to promote C→T (or G→A conversion on the complementary strand) in

a constrained window around the gRNA binding site—allowing for targeted point changes without

extensive DNA fragmentation. This approach was published by Komor et al. in 2016 and paved the

way for point edits with fewer by-products than classic CRISPR/Cas9 (Komor et al., 2016)which can

frequently generate random insertion or deletion of bases (indels.

The extension of the BE repertoire for A→G conversions (ABE — Adenine Base Editors)

was achieved by Gaudelli et al. (2017), through the directed evolution of a tRNA-derived adenosine-

ISSN: 2763-5724 / Vol. 05 - n 05 - ano 2025

deaminase adapted to act on DNA when fused to modied Cas9. ABEs and CBEs have shown, over

time, optimized versions (BE2→BE3→BE4/BE4max; variants of ABE with improved activity and

edit window), along with engineering Cas (high-delity variants) to reduce off-targets and Bystander

edits (Gaudelli et al., 2017).

Prime editing, described by Anzalone et al. in 2019, it broadened the scope by allowing

any→any replacements, small insertions, and deletions without DSBs and without separate exogenous

template. The PE system combines a Cas9 nickase with a reverse transcriptase (RT) and uses a

pegRNA (prime editing guide RNA) that contains the sequence to be written and a tail that serves as

a template for the RT, pegRNA guides the nickase to the target site and provides the information to

be incorporated, which is then synthesized by the RT directly at the site. PE has enormous versatility,

but on the other hand it is a larger protein and, initially, less efcient in some sites than BEs, which

led to successive optimizations (PE2/PE3, improvements in pegRNA design, RT engineering and Cas

variants).

From a mechanistic point of view, the crucial differences that guide the choice between BE

and PE are: (i) type of change desired (BE limited to specic transitions — C→T/T→C or A→G/G→A

— while PE allows transitions, transversions, insertions, and deletions); (ii) edit window and context

(BE has edit window determined by deaminase/Cas and often manages Bystander edits on adjacent

bases, while PE offers greater positional accuracy); and (iii) publisher size and delivery requirements

(PE is structurally larger, complicating delivery by limited-capacity vectors). Recent studies continue

to rene specicity, reduce genomic errors, and characterize cellular repairs that modulate outcomes

(e.g., uracil repair pathways and BE mismatch) (Gu et al., 2025).

Preclinical studies and application models in neurological diseases.

The application of BE and PE in neurological diseases focuses primarily on monogenic

diseases caused by point mutations (or small insertions/deletions) and on strategies to mitigate

ISSN: 2763-5724 / Vol. 05 - n 05 - ano 2025

pathological repeats. Examples and relevant preclinical evidence:

Proof-of-concept in patient-derived cell cultures (iPSC-neurons) and in animal models have

demonstrated that BE can reverse loss-of-function mutations by restoring protein expression and

cellular phenotypes. Gene-driven projects such as MECP2 (Rett syndrome), lysosomal disease genes

such as Tay-Sachs, and other nonsense mutations have shown in vitro edits with sufcient efciency

to consider subsequent in vivo studies (Chang et al., 2021).

Prime editing for complex corrections and nonsense mutations, PE has been applied to

repair premature stop codons and reinstall correct coding sequence in cellular models of neurogenetic

diseases; its ability to generate alterations that are not accessible to BE, such as multinucleotide

transversions or insertions, is especially useful for certain mutations found in rare CNS diseases.

Reports from 2024–2025 describe molecular and phenotypic rescues in animal models for specic

neurological conditions (Sousa et al., 2025).

Repeat expansion diseases, base editing approaches have been explored to “stop” pathogenic

repeats (e.g. CAG in Huntington, introducing substitutions that reduce repeat somatic instability, with

preclinical reports of phenotype mitigation in murine models. These interventions aim both to reduce

the production of toxic protein and to stabilize the genome in the face of expansion (Chiba-Falek,

2025b).

In vivo evidence and transactional relevance, recent advances show that both BE and PE

can be delivered to neural tissue with partial restoration of function and behavioral improvement in

animal models, as the 2024–2025 studies report recovery of enzyme activity, motor improvement,

and reduction of paroxysmal episodes in specic models. These studies mark important preclinical

milestones, but often rely on local vaccination (intracerbral injections) or wide-expression vectors to

demonstrate efcacy (Caso and Davies, 2022)achieving targeted genomic change at unprecedented

efciencies with considerable application in laboratory animal research. Despite its ease of use and

wide application, there remain concerns about the precision of this technology and a number of

unpredictable consequences have been reported, mostly resulting from the DNA double-strand break

ISSN: 2763-5724 / Vol. 05 - n 05 - ano 2025

(DSB.

Delivery to the SNC, safety and technical limitations

Delivery to the central nervous system (CNS) is the main practical limitation for BE/PE

therapies. The blood-brain barrier (BBB), post size (especially for PE), the need to target specic cell

types, and the persistence/timing of expression are critical obstacles. Delivery strategies investigated:

AAVs (adeno-associated viruses), widely used in gene therapy for CNS (good neuronal

tropism proles for certain serotypes), but have limited capacity (~4.7 kb): PE often exceeds this

capacity, requiring strategies such as split-intein systems (splitting the editor into two particles), code

compression, or the use of compact promoters. AAVs have low insertional risk compared to integrative

vectors, but they can induce immune responses and have dose/scale implications. 2024–2025 reviews

discuss AAV optimizations and limitations to deliver bulky editors to the brain (Davis et al., 2024)

prime editing guide RNA stability and modulation of DNA repair. The resulting dual-AAV systems,

v1em and v3em PE-AAV, enable therapeutically relevant prime editing in mouse brain (up to 42%

efciency in cortex.

Lipid nanoparticle (LNP) and non-viral vectors, LNPs, which have proven clinical success

in mRNA vaccines, have been adapted for delivery to the CNS by surface modication or via

direct administrations; LNPs can carry editor-coding RNAs (mRNA) or RNPs, avoiding sustained

expression that can increase immune risk. Recent reviews point to advances in the functionalization

of LNPs to cross BBB and improve tropism, but neuronal efciency and persistence are still variable

(Vargas et al., 2024).

Second Guo et al., (2023), predominant concerns include off-target (unwanted genomic

targets), Bystander edits (for BE, editing of adjacent bases within the window), mosaicism (partial

editing between cells of the same tissue), and immune effects against viral components or exogenous

proteins (Cas9, RT). Studies show that BE has a distinct prole of off-targets, including accidental

ISSN: 2763-5724 / Vol. 05 - n 05 - ano 2025

changes in DNA and, in some cases, deaminase-mediated edits in RNA. PE tends to produce fewer

structural undesirables, but can still generate unwanted insertions/deletions and relies heavily on

pegRNA design and host repair response. Modern protocols employ high-delity Cas variants,

optimized gRNA/pegRNA designs, and deep sequencing pipelines to map off-targets (DNA-seq,

RNA-seq, GUIDE-seq, Digenome-seq, etc.)

Additional technical limitations, variable efciency per locus/genome, chromatin context

dependence, need to optimize conditions for specic cell types (postmitotic neurons versus dividing

cells), and regulatory challenges for permanent CNS interventions. Even with robust proofs of

concept, the safety margin required for human interventions is high, requiring extensive genotoxicity,

immunogenicity, and biodistribution evaluation panels (Schep et al., 2024).

Research perspectives, gaps and priorities.

Based on recent literature, the most relevant priorities and gaps to move BE/PE to the

clinic in neurological diseases are: Robust and scalable delivery platforms for the CNS, developing

optimized AAVs, functionalized LNPs, or novel nanostructures capable of crossing BBB with cellular

selectivity (neuronal vs. glial), high efciency, and lower immunogenicity. Engineering of functional

and compact split-protein systems for PE is critical (An et al., 2024).

Better understanding of the cellular determinants of the editing result, investigate how DNA

repair pathways, chromatin status, and cellular metabolism of neurons inuence BE and PE results, to

reduce variability and mosaicism; studies from 2024–2025 show that repair pathways strongly shape

CBE outcome (Gu et al., 2025).

Strict off-target reduction and mapping, standardize sensitive experimental panels (genomics

and transcriptomics) for off-target detection, and Bystander edits and to develop even more specic

enzyme variants (desaminases/RT and high-delity Cas). Recent work proposes prime editors with

minimized genomic errors and new tools to track off-targets (Chauhan, Sharp and Langer, 2025).

ISSN: 2763-5724 / Vol. 05 - n 05 - ano 2025

More predictive translational models, use of human brain organoids, multicellular co-cultures,

animal models with human-like phenotype, and robust dose/time studies to evaluate long-term

efcacy and safety. Inter-species and inter-model heterogeneity still hinders clinical extrapolations

(Schene et al., 2020).

Ethical, regulatory and access aspects, permanent interventions in the brain raise questions

about consent, reversibility, unknown long-term risks and equity in access. Policies and regulatory

frameworks will need to evolve in parallel with scientic evidence to ensure that clinical trials are

conducted in a safe and socially responsible manner (Wiley et al., 2025).

RESULTS

Table 1: Most relevant publications on the use of base editing and prime editing in neurological

diseases.

Author Year Title of the work Key ndings

Sousa et al. 2025 In vivo prime editing rescues

alternating hemiplegia of

childhood in mice

saram prime editing and base editing to correct

mutations in the ATP1A3 gene (which cause AHC)

in human cell models and two murine models of

HCA. The editing was efcient (up to ~48% in

DNA, ~73% in mRNA), restored ATPase activity,

improved motor and cognitive symptoms, reduced

paroxysmal episodes, and signicantly extended

the lives of mice.

Matuszek et al. 2025 Base editing of trinucleotide

repeats that cause

Huntington’s disease and

Friedreich’s ataxia reduces

somatic repeat expansions in

patient cells and in mice

They developed base editors (CBE and ABE) to

introduce disruptions to the repeated trinucleotides

(CAG in HTT, GAA in FXN) — simulating more

stable alleles. In patient cells and in mice, this

approach reduced the somatic expansion of repeats

in the central nervous system.

BenDavid et al. 2024 Emerging Perspectives on

Prime Editor Delivery to the

Brain

Review article that discusses the challenges and

strategies for delivering prime editors to the brain,

especially considering barriers such as the blood-

brain barrier. It points to nanomedicines and

delivery systems (viral and non-viral) as promising

avenues for neurological gene therapy.

ISSN: 2763-5724 / Vol. 05 - n 05 - ano 2025

Source: Authors, 2025.

DISCUSSION

The experimental studies by Sousa et al. (2025) and de Matuszek et al. (2025) cited in the

Table, represent important preclinical milestones. Sousa et al. demonstrate in vivo correction of

variants that cause Alternating Hemiplegia of Childhood (ATP1A3) using prime editing and base

editing with relevant efciencies and functional recovery in murine models. Matuszek et al. apply

base editors to interrupt trinucleotide repeats (CAG/GAA), reducing somatic expansion in patient

cells and in animal models of Huntington and Friedreich. The review by Bem-David et al. (2024)

complements these results by discussing the practical challenges of delivering prime editors to the

brain. These studies, taken together, show two complementary trajectories, such as the directional

therapeutic application for severe point variants (PE/BE for ATP1A3) and the strategic modication

of repetitive elements to attenuate progressive disease processes (BE for repeats).

The efciency levels reported by Sousa et al., according to values in the range of tens of

percent in DNA and even higher in RNA/protein, with phenotypic rescues, are consistent with the

incremental progress observed since the original descriptions of BE and PE.

Komor et al. (2016) and Gaudelli et al. (2017) showed that base editors can generate high local

efciencies without DSBs, and Anzalone et al. (2019) demonstrated the versatility of Prime Editing

for search-and-replace corrections. Recent in vivo results extend these principles where these authors

have established in vitro capabilities and limitations, and Sousa and Matuszek validate therapeutic

applications in nervous tissue, which conrms technological maturation, but with locus and vector-

dependent efciencies that vary greatly between studies. In other words, the 2025 ndings follow the

trajectory observed since 2016–2019, but now with in vivo proofs of concept that were previously

mostly theoretical.

The main practical difference between the classic benchtop studies and those reported in

ISSN: 2763-5724 / Vol. 05 - n 05 - ano 2025

the table is the delivery solution. Komor et al. (2016), Gaudelli et al. (2017) and Anzalone et al.

(2019) established the editors, however, in vivo translation requires delivery to the CNS and here the

recent publications, including the studies in Table 1, which explore varied strategies such as neonatal

intracerebral injection, split-intein AAVs, optimized LNPs or VLPs (virus-like particles).

Reviews and technical reports such as that of BenDavid (2024), containing work on split-

AAV and VLPs, emphasize that the choice of delivery system determines effective efciency, cell

distribution, and immune safety, which explains why high in vitro efciencies can fall in vivo or vary

between brain regions. Thus, the positive ndings of Sousa et al. (2025) and Matuszek et al. (2025)

demonstrate that therapeutic edits in the CNS are feasible, but depend on delivery solutions that still

require optimization for clinical scalability.

Pioneering work has warned of off-targets, bystander edits (especially for BE), and possible

desaminase-mediated RNA edits. Later studies, as well as by the BE/PE authors themselves, invested

in higher-delit y variants and gRNA /pegRNA designs to mitigate these effects. The 2025 publications

brought here continue this line as they report deep sequencing monitoring and phenotypic toxicity

assessments, but also acknowledge limitations such as mosaicism, possible unintended edits, and

immune response to Cas/vehicle remain real concerns.

Compared to methodological reviews and articles on off-target mapping that use tools such

as GUIDE-seq, Digenome-seq, and RNA-seq, recent studies tend to present more complete safety

panels, but still do not provide long-term follow-up time in humans. In short: there is clear progress

in detecting and reducing adverse effects, but the evidence for long-term safety remains incomplete.

Results such as those of Sousa et al. (2025) on ATP1A3 correction with phenotypic

improvement and Matuszek et al. (2025) on reducing somatic expansion represent crucial translational

steps that show that gene edits can produce signicant functional effects on the CNS. However, when

contrasting with the literature that evaluates clinical applicability, and with the limits described by

Anzalone (2019) and reviews on delivery, some gaps need to be lled before large clinical trials.

Cell distribution and heterogeneity of many diseases require correction in multiple cell types

ISSN: 2763-5724 / Vol. 05 - n 05 - ano 2025

such as neurons, astrocytes, microglia, and microglia, and preclinical studies often focus on specic

regions or populations, since extrapolating to the human brain requires demonstrations of wide and

controlled distribution (O’Carroll, Cook and Young, 2020). Therapeutic scale and window requiring

neonatal corrections or early injections in murine models may not reect efcacy in adult patients

or in diseases with a narrow therapeutic window. This is remembered in reviews on translationality

(Bunuales et al., 2024). Long-term follow-up data on persistence of editing, phenotypic stability, and

late risk of tumorigenesis and cellular dysfunction need long-term studies in large models before

human trials.

As the data in Table 1 t the general picture and methodological recommendations, they

show positive convergence with the ndings that conrm that BE/PE are no longer only in vitro tools,

but also work in vivo in the CNS with measurable therapeutic effects, endorsing the predictions made

by Komor et al. (2025), Gaudelli et al. (2025), and Anzalone et al. (2025). This strengthens the case

for regulatory advances and robust preclinical phase designs.

Regarding the need for an experimental standard, it is recommended that future studies

publish standardized sets of safety data, such as databases of off-targets detected by multiple

techniques, biodistribution by qPCR/sequencing and immune response and make direct comparisons

between vectors and protocols, such as AAV split vs LNP Vs VLP), which is also suggested in the

reviews of BenDavid (2024) and Kalter et al. (2025).

Research priorities are optimization of delivery to the CNS, engineering editors with less

RNA activity, systematic study of mosaicism, and investigation in non-rodent models, such as non-

human primates (when warranted) before clinical transition. These priorities already appear in both

reviews and experimental articles.

CONCLUSION

The consolidation of base editing and prime editing technologies represents a signicant

ISSN: 2763-5724 / Vol. 05 - n 05 - ano 2025

advance in the eld of gene therapy applied to neurological diseases, offering tools capable of correcting

mutations with high precision and minimal generation of double-strand breaks. The analyzed studies

demonstrate that these platforms have already gone beyond the conceptual stage, achieving robust

results in cellular and animal models for pathologies such as Alternating Hemiplegia of Childhood,

Huntington’s disease, and hereditary ataxias. The ability to modulate trinucleotide repeats, correct

pathogenic variants, and restore complex cellular functions in the central nervous system indicates

that next-generation editing systems have real potential for therapeutic interventions in conditions

previously without effective treatment options. These advances reinforce the translational relevance

of CRISPR-based tools and demonstrate that their continuous optimization can redene the landscape

of precision neurological medicine.

However, despite the promising progress, the transition of these technologies to clinical

applications requires caution and systematic investigation of critical aspects still pending. Challenges

include efcient and safe delivery to the CNS, mitigating risks associated with off-target edits and

mosaicism, and the need for long-term safety evidence in more complex preclinical models. In

addition, improvements in vectors, non-viral strategies, and higher-delity editor engineering are

essential to expand clinical applicability. Thus, although base editors and prime editors have already

demonstrated therapeutic feasibility, progress towards clinical practice depends on coordinated efforts

that integrate technological innovation, rigorous functional validation, and biosafety assessment. As a

result, these platforms emerge as central pillars for the future of neurological gene therapies, as long

as they are supported by a solid, continuous and multidisciplinary research agenda.

REFERENCES

A COMPREHENSIVE REVIEW OF AAV-MEDIATED STRATEGIES TARGETING MICROGLIA

FOR THERAPEUTIC INTERVENTION OF NEURODEGENERATIVE DISEASES | JOURNAL

OF NEUROINFLAMMATION. Available at: https://link.springer.com/article/10.1186/s12974-024-

03232-2. Accessed on: 25 nov. 2025.

ISSN: 2763-5724 / Vol. 05 - n 05 - ano 2025

AN, Meirui; RAGURAM, Aditya; DU, Samuel W.; BANSKOTA, Samagya; DAVIS, Jessie R.;

NEWBY, Gregory A.; CHEN, Paul Z.; PALCZEWSKI, Krzysztof; LIU, David R. Engineered virus-

like particles for transient delivery of prime editor ribonucleoprotein complexes in vivo. Nature

Biotechnology, [N.p.], vol. 42, no. 10, p. 1526–1537, out. 2024. https://doi.org/10.1038/s41587-023-

02078-y.

ANZALONE, Andrew V.; RANDOLPH, Peyton B.; DAVIS, Jessie R.; SOUSA, Alexander A.;

KOBLAN, Luke W.; LEVY, Jonathan M.; CHEN, Peter J.; WILSON, Christopher; NEWBY, Gregory

A.; RAGURAM, Aditya; LIU, David R. Search-and-replace genome editing without double-strand

breaks or donor DNA. Nature, vol. 576, no 7785, p. 149–157, Dec. 2019. https://doi.org/10.1038/s41586-

019-1711-4.

BUNUALES, María; GARDUNO, Angeles; CHILLON, Miguel; BOSCH, Assumpció; GONZALEZ-

APARICIO, Manuela; ESPELOSIN, Maria; GARCIA-GOMARA, Marta; RICO, Alberto J.; GARCIA-

OSTA, Ana; CUADRADO-TEJEDOR, Mar; LANCIEGO, José L.; HERNANDEZ-ALCOCEBA,

Ruben. Characterization of brain transduction capability of a BBB-penetrant AAV vector in mice, rats

and macaques reveals differences in expression proles. Gene Therapy, [N.p.], vol. 31, no. 9–10, pp.

455–466, Sept. 2024. https://doi.org/10.1038/s41434-024-00466-w.

CASO, Federico; DAVIES, Benjamin. Base editing and prime editing in laboratory animals.

Laboratory Animals, vol. 56, no. 1, p. 35–49, Feb. 2022. https://doi.org/10.1177/0023677221993895.

CHANG, Kuo-Hsuan; HUANG, Cheng-Yen; OU-YANG, Chih-Hsin; HO, Chang-Han; LIN, Han-Yi;

HSU, Chia-Lang; CHEN, You-Tzung; CHOU, Yu-Chi; CHEN, Yi-Jing; CHEN, Ying; LIN, Jia-Li;

WANG, Ji-Kuan; LIN, Pei-Wen; LIN, Ying-Ru; LIN, Miao-Hsia; TSENG, Chi-Kang; LIN, Chin-

Hsien. In vitro genome editing rescues parkinsonism phenotypes in induced pluripotent stem cells-

derived dopaminergic neurons carrying LRRK2 p.G2019S mutation. Stem Cell Research & Therapy,

Vol. 12, No. 1, p. 508, Sept. 22. 2021. https://doi.org/10.1186/s13287-021-02585-2.

CHAUHAN, Vikash P.; SHARP, Phillip A.; LANGER, Robert. Engineered prime editors with

minimal genomic errors. Nature, Vol. 646, No. 8087, p. 1254–1260, Oct. 2025. https://doi.org/10.1038/

s41586-025-09537-3.

DAVIS, Jessie R.; BANSKOTA, Samagya; LEVY, Jonathan M.; NEWBY, Gregory A.; WANG, Xiao;

ISSN: 2763-5724 / Vol. 05 - n 05 - ano 2025

ANZALONE, Andrew V.; NELSON, Andrew T.; CHEN, Peter J.; HENNES, Andrew D.; AN, Meirui;

ROH, Heejin; RANDOLPH, Peyton B.; MUSUNURU, Kiran; LIU, David R. Efcient prime editing

in mouse brain, liver and heart with dual AAVs. Nature Biotechnology, vol. 42, no. 2, p. 253–264, Feb.

2024. https://doi.org/10.1038/s41587-023-01758-z.

DEVINSKY, Orrin; COLLER, Jeff; AHRENS-NICKLAS, Rebecca; LIU, X. Shawn; AHITUV,

Nadav; DAVIDSON, Beverly L.; BISHOP, Kathie M.; WEISS, Yael; MINGORANCE, Ana. Gene

therapies for neurogenetic disorders. Trends in Molecular Medicine, vol. 31, no. 9, p. 814–826, 1 Sept.

2025. https://doi.org/10.1016/j.molmed.2025.01.015.

FEIGIN, Valery L; VOS, Theo; NICHOLS, Emma; OWOLABI, Mayowa O; CARROLL, William

M; DICHGANS, Martin; DEUSCHL, Günther; PARMAR, Priya; BRAININ, Michael; MURRAY,

Christopher. The global burden of neurological disorders: translating evidence into policy. The Lancet.

Neurology, vol. 19, no. 3, p. 255–265, mar. 2020. https://doi.org/10.1016/S1474-4422(19)30411-9.

GAUDELLI, Nicole M.; KOMOR, Alexis C.; REES, Holly A.; PACKER, Michael S.; BADRAN,

Ahmed H.; BRYSON, David I.; LIU, David R. Programmable base editing of A•T to G•C in genomic

DNA without DNA cleavage. Nature, Vol. 551, No. 7681, pp. 464–471, Nov. 2017. 2017. https://doi.

org/10.1038/nature24644.

GU, Sifeng; BODAI, Zsolt; ANDERSON, Rachel A.; SO, Hei Yu Annika; COWAN, Quinn T.;

KOMOR, Alexis C. Elucidating the genetic mechanisms governing cytosine base editing outcomes

through CRISPRi screens. Nature Communications, [N.p.], vol. 16, no. 1, p. 4685, 20 May 2025a.

https://doi.org/10.1038/s41467-025-59948-z.

GU, Sifeng; BODAI, Zsolt; ANDERSON, Rachel A.; SO, Hei Yu Annika; COWAN, Quinn T.;

KOMOR, Alexis C. Elucidating the genetic mechanisms governing cytosine base editing outcomes

through CRISPRi screens. Nature Communications, [N.p.], vol. 16, no. 1, p. 4685, 20 May 2025b.

https://doi.org/10.1038/s41467-025-59948-z.

GUO, Congting; MA, Xiaoteng; GAO, Fei; GUO, Yuxuan. Off-target effects in CRISPR/Cas9 gene

editing. Frontiers in Bioengineering and Biotechnology, [N.p.], vol. 11, p. 1143157, 9 mar. 2023. https://

doi.org/10.3389/fbioe.2023.1143157.

KALTER, Nechama; FUSTER-GARCÍA, Carla; SILVA, Alfredo; RONCO-DÍAZ, Víctor; RONCELLI,

ISSN: 2763-5724 / Vol. 05 - n 05 - ano 2025

Stefano; TURCHIANO, Giandomenico; GORODKIN, Jan; CATHOMEN, Toni; BENABDELLAH,

Karim; LEE, Ciaran; HENDEL, Ayal. Off-target effects in CRISPR-Cas genome editing for human

therapeutics: Progress and challenges. Molecular Therapy Nucleic Acids, [N.p.], vol. 36, no 3, p.

102636, 9 Sept. 2025. https://doi.org/10.1016/j.omtn.2025.102636.

KANTOR, Ariel; MCCLEMENTS, Michelle E.; MACLAREN, Robert E. CRISPR-Cas9 DNA Base-

Editing and Prime-Editing. International Journal of Molecular Sciences, Vol. 21, No. 17, p. 6240, Aug.

28. 2020. https://doi.org/10.3390/ijms21176240.

KANTOR, Boris; O’DONOVAN, Bernadette; CHIBA-FALEK, Ornit. Trends and challenges of

AAV-delivered gene editing therapeutics for CNS disorders: Implications for neurodegenerative

disease. Molecular Therapy Nucleic Acids, [N.p.], vol. 36, no 3, p. 102635, Sept. 2025a. https://doi.

org/10.1016/j.omtn.2025.102635.

KANTOR, Boris; O’DONOVAN, Bernadette; CHIBA-FALEK, Ornit. Trends and challenges of

AAV-delivered gene editing therapeutics for CNS disorders: Implications for neurodegenerative

disease. Molecular Therapy. Nucleic Acids, [N.p.], vol. 36, no 3, p. 102635, 9 Sept. 2025b. https://doi.

org/10.1016/j.omtn.2025.102635.

KOMOR, Alexis C.; KIM, Yongjoo B.; PACKER, Michael S.; ZURIS, John A.; LIU, David R.

Programmable editing of a target base in genomic DNA without double-stranded DNA cleavage.

Nature, vol. 533, no 7603, p. 420–424 (May 2016). https://doi.org/10.1038/nature17946.

MURRAY, Joss B.; HARRISON, Patrick T.; SCHOLEFIELD, Janine. Prime editing: therapeutic

advances and mechanistic insights. Gene Therapy, [N.p.], vol. 32, no. 2, p. 83–92, mar. 2025. https://

doi.org/10.1038/s41434-024-00499-1.

O’CARROLL, Simon J.; COOK, William H.; YOUNG, Deborah. AAV Targeting of Glial Cell Types

in the Central and Peripheral Nervous System and Relevance to Human Gene Therapy. Frontiers in

Molecular Neuroscience, [N.p.], vol. 13, p. 618020, 2020. https://doi.org/10.3389/fnmol.2020.618020.

PAUL, Abhik; COLLINS, Michael G.; LEE, Hye Young. Gene Therapy: The Next-Generation

Therapeutics and Their Delivery Approaches for Neurological Disorders. Frontiers in Genome Editing,

[N.p.], vol. 4, 22 June 2022. DOI: 10.3389/fgeed.2022.899209. Available at: https://www.frontiersin.

org/journals/genome-editing/articles/10.3389/fgeed.2022.899209/full. Accessed on: 24 nov. 2025.

ISSN: 2763-5724 / Vol. 05 - n 05 - ano 2025

SCHENE, Imre F.; JOORE, Indi P.; OKA, Rurika; MOKRY, Michal; VAN VUGT, Anke H. M.;

VAN BOXTEL, Ruben; VAN DER DOEF, Hubert P. J.; VAN DER LAAN, Luc J. W.; VERSTEGEN,

Monique M. A.; VAN HASSELT, Peter M.; NIEUWENHUIS, Edward E. S.; FUCHS, Sabine A.

Prime editing for functional repair in patient-derived disease models. Nature Communications, [N.p.],

vol. 11, no. 1, p. 5352, 23 Oct. 2020. https://doi.org/10.1038/s41467-020-19136-7.

SOUSA, Alexander A.; TERREY, Markus; SAKAI, Holt A.; SIMMONS, Christine Q.;

ARYSTARKHOVA, Elena; MORSCI, Natalia S.; ANDERSON, Laura C.; XIE, Jun; SURI-PAYER,

Fabian; LAUX, Linda C.; ROZE, Emmanuel; FORLANI, Sylvie; GAO, Guangping; FROST, Simon;

FROST, Nina; SWEADNER, Kathleen J.; GEORGE, Alfred L.; LUTZ, Cathleen M.; LIU, David R.

In vivo prime editing rescues alternating hemiplegia of childhood in mice. Cell, [N.p.], vol. 188, no

16, p. 4275-4294.e23, 7 ago. 2025. https://doi.org/10.1016/j.cell.2025.06.038.

VARGAS, Ronny; LIZANO-BARRANTES, Catalina; ROMERO, Miquel; VALENCIA-CLUA,

Kevin; NARVÁEZ-NARVÁEZ, David A.; SUÑÉ-NEGRE, Josep Ma; PÉREZ-LOZANO, Pilar;

GARCÍA-MONTOYA, Encarna; MARTINEZ-MARTINEZ, Noelia; HERNÁNDEZ-MUNAIN,

Cristina; SUÑÉ, Carlos; SUÑÉ-POU, Marc. The piper at the gates of brain: A systematic review

of surface modication strategies on lipid nanoparticles to overcome the Blood-Brain-Barrier.

International Journal of Pharmaceutics, [N.p.], vol. 665, p. 124686, 15 Nov. 2024. https://doi.

org/10.1016/j.ijpharm.2024.124686.

WILEY, Lindsay; CHEEK, Mattison; LAFAR, Emily; MA, Xiaolu; SEKOWSKI, Justin;

TANGUTURI, Nikki; ILTIS, Ana. The Ethics of Human Embryo Editing via CRISPR-Cas9

Technology: A Systematic Review of Ethical Arguments, Reasons, and Concerns. Hec Forum, [N.p.],

vol. 37, no 2, p. 267–303, 2025. https://doi.org/10.1007/s10730-024-09538-1.