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

作　　者：Shibo Hou Jingheng Chen Ruobing Feng Xuejiao Xu Nan Liang Jackson Champer 

机构地区：[1]Center for Bioinformatics,Center for Life Sciences,School of Life Sciences,Peking University,Beijing 100871,China

出　　处：《Journal of Genetics and Genomics》2024年第8期836-843,共8页遗传学报（英文版）

基　　金：supported by laboratory startup funds from Peking University and the Center for Life Sciences,as well as the grants from the National Science Foundation of China(32302455 and 32270672)。

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 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.

关 键 词：Genedrive Homing drive Rescueelement Population modification Multiplexed gRNAs Resistance alleles Cage study 

分 类 号：R744.51[医药卫生—神经病学与精神病学]