CRISPR-KRISPR: a method to identify on-target and random insertion of donor DNAs and their characterization in knock-in mice.

CRISPR-KRISPR: a method to identify on-target and random insertion of donor DNAs and their characterization in knock-in mice.
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DOI:
10.1186/s13059-022-02779-8
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发表时间:
2022-10-25
期刊:
影响因子:
12.3
通讯作者:
--
中科院分区:
生物学1区
文献类型:
--
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CRISPR工具可以很容易地生成敲除和敲入动物模型,但这些模型可能包含脱靶基因组损伤或供体DNA的随机插入。更简单的方法来识别脱靶病变和随机插入,使用尾部或耳端DNA,是不可用的。我们开发了CRISPR-KRISPR(CRISPR-Knock-ins and Random Sequence Searching PRotocol),这是一种识别脱靶病变和随机插入的方法。CRISPR-KRISPR仅使用3.4 μg的基因组DNA;因此,它可以很容易地作为一个额外的步骤纳入基因型建立者动物进一步育种。在线版本包含补充材料,可通过10.1186/s13059-022-02779-8获得。
CRISPR tools can generate knockout and knock-in animal models easily, but the models can contain off-target genomic lesions or random insertions of donor DNAs. Simpler methods to identify off-target lesions and random insertions, using tail or earpiece DNA, are unavailable. We develop CRISPR-KRISPR (CRISPR-Knock-ins and Random Inserts Searching PRotocol), a method to identify both off-target lesions and random insertions. CRISPR-KRISPR uses as little as 3.4 μg of genomic DNA; thus, it can be easily incorporated as an additional step to genotype founder animals for further breeding. The online version contains supplementary material available at 10.1186/s13059-022-02779-8.
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