期刊
MOLECULAR CELL
卷 80, 期 2, 页码 246-+出版社
CELL PRESS
DOI: 10.1016/j.molcel.2020.09.003
关键词
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资金
- NIH [R01 GM117321, DP5OD023098]
- Paul G. Allen Frontiers Group Distinguished Investigator Award
- Safe Genes program of the Defense Advanced Research Projects Agency (DARPA) [HR0011-17-2-0047]
CRISPR-Cas9-based gene drive systems possess the inherent capacity to spread progressively throughout target populations. Here we describe two self-copying (or active) guide RNA-only genetic elements, called e-CHACRs and ERACRs. These elements use Cas9 produced in trans by a gene drive either to inactivate the cas9 transgene (e-CHACRs) or to delete and replace the gene drive (ERACRs). e-CHACRs can be inserted at various genomic locations and carry two or more gRNAs, the first copying the e-CHACR and the second mutating and inactivating the cas9 transgene. Alternatively, ERACRs are inserted at the same genomic location as a gene drive, carrying two gRNAs that cut on either side of the gene drive to excise it. e-CHACRs efficiently inactivate Cas9 and can drive to completion in cage experiments. Similarly, ERACRs, particularly those carrying a recoded cDNA-restoring endogenous gene activity, can drive reliably to fully replace a gene drive. We compare the strengths of these two systems.
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