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CATI: an efficient gene integration method for rodent and primate embryos by MMEJ suppression

The efficiency of homology-directed repair (HDR) plays a crucial role in the development of animal models and gene therapy. We demonstrate that microhomology-mediated end-joining (MMEJ) constitutes a substantial proportion of DNA repair during CRISPR-mediated gene editing. Using CasRx to downregulat...

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Detalles Bibliográficos
Autores principales: Chen, Hongyu, Liu, Xingchen, Li, Lanxin, Tan, Qingtong, Li, Shiyan, Li, Li, Li, Chunyang, Fu, Jiqiang, Lu, Yong, Wang, Yan, Sun, Yidi, Luo, Zhen-Ge, Lu, Zongyang, Sun, Qiang, Liu, Zhen
Formato: Online Artículo Texto
Lenguaje:English
Publicado: BioMed Central 2023
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC10288798/
https://www.ncbi.nlm.nih.gov/pubmed/37353834
http://dx.doi.org/10.1186/s13059-023-02987-w
Descripción
Sumario:The efficiency of homology-directed repair (HDR) plays a crucial role in the development of animal models and gene therapy. We demonstrate that microhomology-mediated end-joining (MMEJ) constitutes a substantial proportion of DNA repair during CRISPR-mediated gene editing. Using CasRx to downregulate a key MMEJ factor, Polymerase Q (Polq), we improve the targeted integration efficiency of linearized DNA fragments and single-strand oligonucleotides (ssODN) in mouse embryos and offspring. CasRX-assisted targeted integration (CATI) also leads to substantial improvements in HDR efficiency during the CRISPR/Cas9 editing of monkey embryos. We present a promising tool for generating monkey models and developing gene therapies for clinical trials. SUPPLEMENTARY INFORMATION: The online version contains supplementary material available at 10.1186/s13059-023-02987-w.