Methods for improving crisper / cas9 insertion efficiency in non-human mammalian embryos

By using the compound AZD7648 and the RNA editor CasRX that inhibits Polq expression, the MMEJ repair pathway was blocked, and HDR repair was promoted, thus solving the problem of low CRISPR/Cas9 gene knock-in efficiency in non-human mammalian embryos and achieving highly efficient gene editing.

CN122405745APending Publication Date: 2026-07-17CENT FOR EXCELLENCE IN BRAIN SCI & INTELLIGENCE TECH CHINESE ACAD OF SCI
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Patent Information

Application Number
CN202510073394.1
Authority / Receiving Office
CN · China
Patent Type
Applications(China)
Current Assignee / Owner
Filing Date
2025-01-16
Publication Date
2026-07-17

AI Technical Summary

Technical Problem

In non-human mammalian embryos, the efficiency of gene knock-in using the CRISPR/Cas9 system is low, especially when drug screening or flow cytometry sorting is not possible. Existing methods make it difficult to select suitable sgRNAs to achieve efficient gene knock-in.

Method used

Using the compound AZD7648 and the RNA editor CasRX that inhibits Polq expression, we can improve the gene editing efficiency of CRISPR/Cas9 by blocking the MMEJ repair pathway and promoting HDR repair. This method is called CCEH.

Benefits of technology

It significantly improves the gene knock-in efficiency of sgRNA targets, up to 90%, and provides a universal and efficient gene editing method applicable to non-human mammalian embryos.

✦ Generated by Eureka AI based on patent content.

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Abstract

本发明提供了一种提高CRISPR / Cas9在非人哺乳动物胚胎中插入效率的方法，包括使用式(I)所示的化合物和抑制Polq表达的RNA编辑器CasRX对非人类哺乳动物的胚胎进行处理，其中式(I)所示的化合物包括式(I)所示的化合物，或其药学上可接受的盐。
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