Next-Generation CRISPR Technologies and Their Applications in Gene and Cell Therapy

M Alejandra Zeballos C; Thomas Gaj

doi:10.1016/j.tibtech.2020.10.010

Next-Generation CRISPR Technologies and Their Applications in Gene and Cell Therapy

Trends Biotechnol. 2021 Jul;39(7):692-705. doi: 10.1016/j.tibtech.2020.10.010.

Authors

M Alejandra Zeballos C¹, Thomas Gaj²

Affiliations

¹ Department of Bioengineering, University of Illinois, Urbana, IL 61801, USA.
² Department of Bioengineering, University of Illinois, Urbana, IL 61801, USA; Carl R. Woese Institute for Genomic Biology, University of Illinois, Urbana, IL 61801, USA. Electronic address: gaj@illinois.edu.

Abstract

The emergence of clustered regularly interspaced short palindromic repeat (CRISPR) nucleases has transformed biotechnology by providing an easy, efficient, and versatile platform for editing DNA. However, traditional CRISPR-based technologies initiate editing by activating DNA double-strand break (DSB) repair pathways, which can cause adverse effects in cells and restrict certain therapeutic applications of the technology. To this end, several new CRISPR-based modalities have been developed that are capable of catalyzing editing without the requirement for a DSB. Here, we review three of these technologies: base editors, prime editors, and RNA-targeting CRISPR-associated protein (Cas)13 effectors. We discuss their strengths compared to traditional gene-modifying systems, we highlight their emerging therapeutic applications, and we examine challenges facing their safe and effective clinical implementation.

Keywords: CRISPR; CRISPR-Cas13; base editing; gene therapy; prime editing.

Publication types

Research Support, N.I.H., Extramural
Research Support, Non-U.S. Gov't
Review

MeSH terms

CRISPR-Cas Systems*
Cell- and Tissue-Based Therapy / trends
Endonucleases / genetics
Gene Editing*
Humans

Substances

Endonucleases

Grants and funding

T32 EB019944/EB/NIBIB NIH HHS/United States