A homing rescue gene drive with multiplexed gRNAs reaches high frequency in cage populations but generates functional resistance.

Autor: Hou S; Center for Bioinformatics, Center for Life Sciences, School of Life Sciences, Peking University, Beijing 100871, China., Chen J; Center for Bioinformatics, Center for Life Sciences, School of Life Sciences, Peking University, Beijing 100871, China., Feng R; Center for Bioinformatics, Center for Life Sciences, School of Life Sciences, Peking University, Beijing 100871, China., Xu X; Center for Bioinformatics, Center for Life Sciences, School of Life Sciences, Peking University, Beijing 100871, China., Liang N; Center for Bioinformatics, Center for Life Sciences, School of Life Sciences, Peking University, Beijing 100871, China., Champer J; Center for Bioinformatics, Center for Life Sciences, School of Life Sciences, Peking University, Beijing 100871, China. Electronic address: jchamper@pku.edu.cn.
Jazyk: angličtina
Zdroj: Journal of genetics and genomics = Yi chuan xue bao [J Genet Genomics] 2024 Aug; Vol. 51 (8), pp. 836-843. Date of Electronic Publication: 2024 Apr 08.
DOI: 10.1016/j.jgg.2024.04.001
Abstrakt: CRISPR homing gene drives have considerable potential for managing populations of medically and agriculturally significant insects. They operate by Cas9 cleavage followed by homology-directed repair, copying the drive allele to the wild-type chromosome and thus increasing in frequency and spreading throughout a population. However, resistance alleles formed by end-joining repair pose a significant obstacle. To address this, we create a homing drive targeting the essential hairy gene in Drosophila melanogaster. Nonfunctional resistance alleles are recessive lethal, while drive carriers have a recoded "rescue" version of hairy. The drive inheritance rate is moderate, and multigenerational cage studies show drive spread to 96%-97% of the population. However, the drive does not reach 100% due to the formation of functional resistance alleles despite using four gRNAs. These alleles have a large deletion but likely utilize an alternate start codon. Thus, revised designs targeting more essential regions of a gene may be necessary to avoid such functional resistance. Replacement of the rescue element's native 3' UTR with a homolog from another species increases drive inheritance by 13%-24%. This was possibly because of reduced homology between the rescue element and surrounding genomic DNA, which could also be an important design consideration for rescue gene drives.
Competing Interests: Conflict of interest The authors declare that they have no conflict of interest.
(Copyright © 2024 Institute of Genetics and Developmental Biology, Chinese Academy of Sciences, and Genetics Society of China. Published by Elsevier Ltd. All rights reserved.)
Databáze: MEDLINE
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