# Genetic foundation for complete mutant allele-specific CRISPR in neurodegenerative diseases

> **NIH NIH R01** · MASSACHUSETTS GENERAL HOSPITAL · 2021 · $456,205

## Abstract

SUMMARY / ABSTRACT
Title: Genetic foundation for complete mutant allele-specific CRISPR in neurodegenerative diseases
Many cases of dominant neurodegenerative disorders defy the development of effective treatments despite
their long-known genetic causes and numerous implicated underlying pathways, reflecting difficulty in defining
rational drug targets through investigations focusing on mechanisms. Alternatively, development and
maturation of gene targeting/lowering approaches have highlighted the value of disease-causing mutation itself
as the target of treatments. Although promising, mRNA-lowering approaches suffer from inherent limitations of
requirement of repeated treatments, allele-specificity, and dosage controls. We recently developed a complete
allele-specific DNA targeting strategy based on CRISPR gene editing technology using PAM-altering SNP to
overcome key limitations of conventional mRNA targeting approaches. Our silencing strategy achieves perfect
allele specificity by using SNP variations that create CRISPR PAM sites selectively on the mutant
allele/haplotype. Importantly, our novel CRISPR gene silencing strategy targets the haplotype backbone that
carries the disease-causing mutation, and therefore does not depend on the type, size, and location of the
disease allele, providing broadly applicable therapeutic platforms. Here, we propose to determine therapeutic
potential of PAM-Altering SNP (PAS)-based CRISPR strategies 1) to prevent the transcription or 2) to induce
nonsense-mediated decay of the mutant allele in cells and animal models of Huntington's disease (HD).
Briefly, we will: 1) determine whether mutant allele-specific Transcription Prevention by CRISPR (TP-CRISPR)
and Nonsense-Mediated Decay by CRISPR (NMD-CRISPR) efficiently introduce knockout mutations in
neuronal cells derived from patients and brains of animal models of HD, 2) test whether selective silencing of
mutant allele of a developmentally important gene (i.e., HTT) influences neuronal differentiation capability of
induced pluripotent stem cells (iPSC), 3) compare targeting efficiency in neurons and glia, and 4) determine
molecular/cellular, behavioral consequences, and pre-clinical therapeutic efficacy of mutant allele-specific
CRISPR in vivo. We anticipate this research will 1) nominate optimal CRISPR targeting sites and strategies for
HD, 2) generate knowledge base regarding efficiency of mutant-specific CRISPR, and 3) evaluate potential of
PAS-based allele-specific CRISPR as therapeutic intervention for HD, providing 1) genetic foundation for novel
and innovative therapeutic routes for HD and 2) proof-of-concept for other dominant neurodegenerative
diseases.

## Key facts

- **NIH application ID:** 10216366
- **Project number:** 5R01NS105709-04
- **Recipient organization:** MASSACHUSETTS GENERAL HOSPITAL
- **Principal Investigator:** Jong-Min Lee
- **Activity code:** R01 (R01, R21, SBIR, etc.)
- **Funding institute:** NIH
- **Fiscal year:** 2021
- **Award amount:** $456,205
- **Award type:** 5
- **Project period:** 2018-09-15 → 2023-06-30

## Primary source

NIH RePORTER: https://reporter.nih.gov/project-details/10216366

## Citation

> US National Institutes of Health, RePORTER application 10216366, Genetic foundation for complete mutant allele-specific CRISPR in neurodegenerative diseases (5R01NS105709-04). Retrieved via AI Analytics 2026-05-22 from https://api.ai-analytics.org/grant/nih/10216366. Licensed CC0.

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