Engineered Cas12i2 is a Versatile High-efficiency Platform for Therapeutic Genome Editing

Overview

Journal Nat Commun

Specialty Biology

Date 2022 May 20

PMID 35595757

Authors

Colin McGaw

Anthony J Garrity

Gabrielle Z Munoz

Jeffrey R Haswell

Sejuti Sengupta

Elise Keston-Smith

Pratyusha Hunnewell

Alexa Ornstein

Mishti Bose

Quinton Wessells

Noah Jakimo

Paul Yan

Huaibin Zhang

Lauren E Alfonse

Roy Ziblat

Jason M Carte

Wei-Cheng Lu

Derek Cerchione

Brendan Hilbert

Shanmugapriya Sothiselvam

Winston X Yan

David R Cheng

David A Scott

Tia DiTommaso

Shaorong Chong

Affiliations

Soon will be listed here.

Abstract

The CRISPR-Cas type V-I is a family of Cas12i-containing programmable nuclease systems guided by a short crRNA without requirement for a tracrRNA. Here we present an engineered Type V-I CRISPR system (Cas12i), ABR-001, which utilizes a tracr-less guide RNA. The compact Cas12i effector is capable of self-processing pre-crRNA and cleaving dsDNA targets, which facilitates versatile delivery options and multiplexing, respectively. We apply an unbiased mutational scanning approach to enhance initially low editing activity of Cas12i2. The engineered variant, ABR-001, exhibits broad genome editing capability in human cell lines, primary T cells, and CD34+ hematopoietic stem and progenitor cells, with both robust efficiency and high specificity. In addition, ABR-001 achieves a high level of genome editing when delivered via AAV vector to HEK293T cells. This work establishes ABR-001 as a versatile, specific, and high-performance platform for ex vivo and in vivo gene therapy.

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