# Engineering of a mouse model of choline acetycholinesterase deficients using CRISPR/Cas9 gene editing

> **NIH NIH R03** · UNIVERSITY OF CALIFORNIA AT DAVIS · 2022 · $159,750

## Abstract

ENGINEERING OF A MOUSE MODEL OF CHOLINE ACETYLTRANSFERASE DEFICIENCY USING
CRISPR/CAS9 GENE EDITING
Scientific Abstract:
Choline acetyltransferase (ChAT) is the enzyme that resynthesizes the neurotransmitter acetylcholine, from
acetyl-CoA and choline at the soma of multiple types of neurons of the peripheral and central nervous system
(CNS). Mutations in the ChAT gene (CHAT) result in a potentially fatal form of human congenital myasthenic
syndrome (CMS) referred to as CMS associated with episodic apneas. Chat knockout (KO) mice are immobile
and die at birth, and conditional Chat KO (Chatflox/flox/Cre-ERT2) mice die a few weeks after the induction of Chat
ablation by an intraperitoneal injection of tamoxifen. However, the lethal Chatflox/flox/Cre-ERT2 phenotype can be
rescued in most cases by a single intracranial injection at the first day of life of 2x1013 vg/kg adeno-associated
virus type 9 (AAV9) carrying the human CHAT gene (AAV9-CHAT). Furthermore, injected mice can reach adult
life without developing obvious weakness. While these results are encouraging, the widespread depletion of
Chat induced in the conditional knock out model is not an adequate model of the human disease, nor the
injection of the therapeutic agent at the first day of life reflects the most likely time of therapeutic interventions
in humans. Thus, to refine the model of CMS due to ChAT deficiency, we plan to create knock in (KI) mice
homozygous for two previously identified human CHAT mutations, p.Val136 Met and p.Arg207His, using
CRISPR/Cas9 gene editing. We will then attempt to correct the phenotype of ChatKI/KI mice by injecting
intracisternally AAV9-CHAT during early postnatal life. The overall goal of the project is to generate an animal
model that will reproduce as faithfully as possible the human disease and to conduct a pilot feasibility study on
the efficacy of AAV9-CHAT mediated gene therapy in this newly developed animal model of CMS caused by
CHAT mutations.

## Key facts

- **NIH application ID:** 10511979
- **Project number:** 1R03NS128810-01
- **Recipient organization:** UNIVERSITY OF CALIFORNIA AT DAVIS
- **Principal Investigator:** RICARDO Anibal MASELLI
- **Activity code:** R03 (R01, R21, SBIR, etc.)
- **Funding institute:** NIH
- **Fiscal year:** 2022
- **Award amount:** $159,750
- **Award type:** 1
- **Project period:** 2022-07-01 → 2024-06-30

## Primary source

NIH RePORTER: https://reporter.nih.gov/project-details/10511979

## Citation

> US National Institutes of Health, RePORTER application 10511979, Engineering of a mouse model of choline acetycholinesterase deficients using CRISPR/Cas9 gene editing (1R03NS128810-01). Retrieved via AI Analytics 2026-06-10 from https://api.ai-analytics.org/grant/nih/10511979. Licensed CC0.

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