Engineering of SauriCas9 with enhanced specificity

Engineering of SauriCas9 with enhanced specificity

SauriCas9 is a compact Cas9 nuclease showing promise for in vivo therapeutic applications. However, concerns about off-target effects necessitated improvements in specificity. We addressed this by introducing mutations to eliminate polar contacts between Cas9 and the target DNA, resulting in the Sau...

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Bibliographic Details
Main Authors: Xiaoqi Zhang, Chen Tao, Miaomiao Li, Sufang Zhang, Puping Liang, Yan Huang, Huihui Liu, Yongming Wang
Format: Article
Language:English
Published: Elsevier 2025-03-01
Series:Molecular Therapy: Nucleic Acids
Subjects:
MT: RNA/DNA Editing
gene editing
CRISPR-Cas
compact Cas9
SauriCas9-R253A
specificity
Online Access:http://www.sciencedirect.com/science/article/pii/S2162253125000095
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Summary:SauriCas9 is a compact Cas9 nuclease showing promise for in vivo therapeutic applications. However, concerns about off-target effects necessitated improvements in specificity. We addressed this by introducing mutations to eliminate polar contacts between Cas9 and the target DNA, resulting in the SauriCas9-R253A variant with enhanced specificity. To validate its efficacy, we employed SauriCas9-R253A to disrupt three genes (B2M, TRAC, and PDCD1), a strategy integral to the development of allogeneic chimeric antigen receptor T cell (CAR-T) therapies. Our results demonstrated that the most efficient single-guide RNAs for SauriCas9-R253A exhibited comparable activity to SpCas9 and showed no detectable off-target effects in the disruption of these genes, highlighting its therapeutic potential.
ISSN:2162-2531

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