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

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外文摘要：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 mel-anogaster.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.

外文关键词：

Gene driveHoming driveRescue elementPopulation modificationMultiplexed gRNAsResistance allelesCage study

作者：

Shibo Hou、Jingheng Chen、Ruobing Feng、Xuejiao Xu、Nan Liang、Jackson Champer

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作者单位：

Center for Bioinformatics,Center for Life Sciences,School of Life Sciences,Peking University,Beijing 100871,China

基金：

laboratory startup funds from Peking University and the Center for Life SciencesNational Science Foundation of ChinaNational Science Foundation of China

项目编号：

3230245532270672

出版年：

2024

DOI：

10.1016/j.jgg.2024.04.001

遗传学报

中国遗传学会中国科学院遗传与发育生物学研究所

遗传学报

CSTPCD

影响因子：0.821

ISSN：1673-8527

年,卷(期)：2024.51(8)