Document Type

Article

Publication Date

6-12-2020

Keywords

JGM, JMG, JAXCC

JAX Source

Nat Commun 2020 Jun 12; 11(1):2973

PMID

32532987

DOI

https://doi.org/10.1038/s41467-020-16806-4

Grant

HG009900,CA034196,Leukemia Research Foundation New Investigator Grant, The Jackson Laboratory Director's Innovation Fund.

Abstract

Alternative splicing allows expression of mRNA isoforms from a single gene, expanding the diversity of the proteome. Its prevalence in normal biological and disease processes warrant precise tools for modulation. Here we report the engineering of CRISPR Artificial Splicing Factors (CASFx) based on RNA-targeting CRISPR-Cas systems. We show that simultaneous exon inclusion and exclusion can be induced at distinct targets by differential positioning of CASFx. We also create inducible CASFx (iCASFx) using the FKBP-FRB chemical-inducible dimerization domain, allowing small molecule control of alternative splicing. Finally, we demonstrate the activation of SMN2 exon 7 splicing in spinal muscular atrophy (SMA) patient fibroblasts, suggesting a potential application of the CASFx system.

Comments

We thank the Jackson Laboratory Flow Cytometry for FACS experiments, and Jackson Laboratory Research Program Development for help with manuscript editing.

This article is licensed under a Creative Commons Attribution 4.0 International License.

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