Targeted Gene Silencing in the Nervous System with CRISPR-Cas13

Overview

Journal Sci Adv

Specialties Biology
Science

Date 2022 Jan 19

PMID 35044815

Authors

Jackson E Powell

Colin K W Lim

Ramya Krishnan

Tristan X McCallister

Christian Saporito-Magrina

Maria A Zeballos

Garrett D McPheron

Thomas Gaj

Affiliations

Soon will be listed here.

Abstract

Cas13 nucleases are a class of programmable RNA-targeting CRISPR effector proteins that are capable of silencing target gene expression in mammalian cells. Here, we demonstrate that RfxCas13d, a Cas13 ortholog with favorable characteristics to other family members, can be delivered to the mouse spinal cord and brain to silence neurodegeneration-associated genes. Intrathecally delivering an adeno-associated virus vector encoding an RfxCas13d variant programmed to target superoxide dismutase 1 (SOD1), a protein whose mutation can cause amyotrophic lateral sclerosis, reduced SOD1 mRNA and protein in the spinal cord by >50% and improved outcomes in a mouse model of the disorder. We further show that intrastriatally delivering an RfxCas13d variant programmed to target huntingtin (HTT), a protein whose mutation is causative for Huntington’s disease, led to a ~50% reduction in HTT protein in the mouse brain. Our results establish RfxCas13d as a versatile platform for knocking down gene expression in the nervous system.

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