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Bridging Gaps in HDR Improvement: The Role of MAD2L2, SCAI, and SCR7.


ABSTRACT: This study aimed to enhance homology-directed repair (HDR) efficiency in CRISPR/Cas-mediated genome editing by targeting three key factors regulating the balance between HDR and non-homologous end joining (NHEJ): MAD2L2, SCAI, and Ligase IV. In order to achieve this, a cellular model using mutated eGFP was designed to monitor HDR events. Results showed that MAD2L2 knockdown and SCR7 treatment significantly improved HDR efficiency during Cas9-mediated HDR repair of the mutated eGFP gene in the HEK293T cell line. Fusion protein Cas9-SCAI did not improve HDR. This study is the first to demonstrate that MAD2L2 knockdown during CRISPR-mediated gene editing in HEK293T cells can increase precise correction by up to 10.2 times. The study also confirmed a moderate but consistent effect of SCR7, an inhibitor of Ligase IV, which increased HDR by 1.7 times. These findings provide valuable insights into improving HDR-based genome editing efficiency.

SUBMITTER: Anuchina AA 

PROVIDER: S-EPMC10095018 | biostudies-literature | 2023 Apr

REPOSITORIES: biostudies-literature

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Bridging Gaps in HDR Improvement: The Role of MAD2L2, SCAI, and SCR7.

Anuchina Arina A AA   Zaynitdinova Milyausha I MI   Demchenko Anna G AG   Evtushenko Nadezhda A NA   Lavrov Alexander V AV   Smirnikhina Svetlana A SA  

International journal of molecular sciences 20230404 7


This study aimed to enhance homology-directed repair (HDR) efficiency in CRISPR/Cas-mediated genome editing by targeting three key factors regulating the balance between HDR and non-homologous end joining (NHEJ): MAD2L2, SCAI, and Ligase IV. In order to achieve this, a cellular model using mutated eGFP was designed to monitor HDR events. Results showed that MAD2L2 knockdown and SCR7 treatment significantly improved HDR efficiency during Cas9-mediated HDR repair of the mutated eGFP gene in the HE  ...[more]

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