A Fluorescent Reporter Mouse for In Vivo Assessment of Genome Editing with Diverse Cas Nucleases and Prime Editors

Zexiang Chen; Suet-Yan Kwan; Aamir Mir; Max Hazeltine; Minwook Shin; Shun-Qing Liang; Io Long Chan; Karen Kelly; Krishna S Ghanta; Nicholas Gaston; Yueying Cao; Jun Xie; Guangping Gao; Wen Xue; Erik J Sontheimer; Jonathan K Watts

doi:10.1089/crispr.2023.0048

A Fluorescent Reporter Mouse for In Vivo Assessment of Genome Editing with Diverse Cas Nucleases and Prime Editors

CRISPR J. 2023 Dec;6(6):570-582. doi: 10.1089/crispr.2023.0048.

Authors

Zexiang Chen¹, Suet-Yan Kwan¹, Aamir Mir¹, Max Hazeltine¹, Minwook Shin¹, Shun-Qing Liang¹, Io Long Chan¹, Karen Kelly¹, Krishna S Ghanta¹, Nicholas Gaston¹, Yueying Cao¹, Jun Xie^{2

3

4

5

6}, Guangping Gao^{2

3

4

5

6}, Wen Xue^{1

2

7

8}, Erik J Sontheimer^{1

2

7

9}, Jonathan K Watts^{1

2

3

9}

Affiliations

¹ RNA Therapeutics Institute, University of Massachusetts Chan Medical School, Worcester, Massachusetts, USA.
² Li Weibo Institute for Rare Diseases Research, University of Massachusetts Chan Medical School, Worcester, Massachusetts, USA.
³ NeuroNexus Institute, University of Massachusetts Chan Medical School, Worcester, Massachusetts, USA.
⁴ Horae Gene Therapy Center, University of Massachusetts Chan Medical School, Worcester, Massachusetts, USA.
⁵ Viral Vector Core, University of Massachusetts Chan Medical School, Worcester, Massachusetts, USA.
⁶ Department of Microbiology and Physiological Systems, University of Massachusetts Chan Medical School, Worcester, Massachusetts, USA.
⁷ Program in Molecular Medicine, University of Massachusetts Chan Medical School, Worcester, Massachusetts, USA.
⁸ Department of Molecular, Cell and Cancer Biology, University of Massachusetts Chan Medical School, Worcester, Massachusetts, USA.
⁹ Department of Biochemistry and Molecular Biotechnology, University of Massachusetts Chan Medical School, Worcester, Massachusetts, USA.

Abstract

CRISPR-based genome-editing technologies, including nuclease editing, base editing, and prime editing, have recently revolutionized the development of therapeutics targeting disease-causing mutations. To advance the assessment and development of genome editing tools, a robust mouse model is valuable, particularly for evaluating in vivo activity and delivery strategies. In this study, we successfully generated a knock-in mouse line carrying the Traffic Light Reporter design known as TLR-multi-Cas variant 1 (TLR-MCV1). We comprehensively validated the functionality of this mouse model for both in vitro and in vivo nuclease and prime editing. The TLR-MCV1 reporter mouse represents a versatile and powerful tool for expediting the development of editing technologies and their therapeutic applications.

MeSH terms

Animals
CRISPR-Cas Systems* / genetics
Disease Models, Animal
Endonucleases / genetics
Gene Editing*
Mice
Technology

Substances

Endonucleases

Abstract

MeSH terms

Substances

Grants and funding