Abstract：
:CRISPR-Cas9 is a powerful new tool for genome editing, but this technique creates mosaic mutations that affect the efficiency and precision of its ability to edit the genome. Reducing mosaic mutations is particularly important for gene therapy and precision genome editing. Although the mechanisms underlying the CRSIPR/Cas9-mediated mosaic mutations remain elusive, the prolonged expression and activity of Cas9 in embryos could contribute to mosaicism in DNA mutations. Here we report that tagging Cas9 with ubiquitin-proteasomal degradation signals can facilitate the degradation of Cas9 in non-human primate embryos. Using embryo-splitting approach, we found that shortening the half-life of Cas9 in fertilized zygotes reduces mosaic mutations and increases its ability to modify genomes in non-human primate embryos. Also, injection of modified Cas9 in one-cell embryos leads to live monkeys with the targeted gene modifications. Our findings suggest that modifying Cas9 activity can be an effective strategy to enhance precision genome editing.
journal_name
Sci Repjournal_title
Scientific reportsauthors
Tu Z,Yang W,Yan S,Yin A,Gao J,Liu X,Zheng Y,Zheng J,Li Z,Yang S,Li S,Guo X,Li XJdoi
10.1038/srep42081subject
Has Abstractpub_date
2017-02-03 00:00:00pages
42081issn
2045-2322pii
srep42081journal_volume
7pub_type
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