phaidra.vetmeduni.ac.at o:1864 DOI 10.1093/g3journal/jkac081 A homing suppression gene drive with multiplexed gRNAs maintains high drive conversion efficiency and avoids functional resistance alleles Gene drives are engineered alleles that can bias inheritance in their favor, allowing them to spread throughout a population. They could potentially be used to modify or suppress pest populations, such as mosquitoes that spread diseases. CRISPR/Cas9 homing drives, which copy themselves by homology-directed repair in drive/wild-type heterozygotes, are a powerful form of gene drive, but they are vulnerable to resistance alleles that preserve the function of their target gene. Such resistance alleles can prevent successful population suppression. Here, we constructed a homing suppression drive in Drosophila melanogaster that utilized multiplexed gRNAs to inhibit the formation of functional resistance alleles in its female fertility target gene. The selected gRNA target sites were close together, preventing reduction in drive conversion efficiency. The construct reached a moderate equilibrium frequency in cage populations without apparent formation of resistance alleles. However, a moderate fitness cost prevented elimination of the cage population, showing the importance of using highly efficient drives in a suppression strategy, even if resistance can be addressed. Nevertheless, our results experimentally demonstrate the viability of the multiplexed gRNAs strategy in homing suppression gene drives. eng Malaria Vector Mosquito; Population Modification; Nucleases; Evolution; Fate 2023-07-19T10:17:16.132Z LOMv1.0 Author BEGIN:VCARD VERSION:3.0 N:Yang;Emily; FN:Emily Yang END:VCARD BEGIN:VCARD VERSION:3.0 N:Champer;Jackson; FN:Jackson Champer X-ORCID:https://orcid.org/0000-0002-3814-3774 END:VCARD BEGIN:VCARD VERSION:3.0 N:Messer;Philipp W.; FN:Philipp W. Messer X-ORCID:https://orcid.org/0000-0001-8453-9377 END:VCARD BEGIN:VCARD VERSION:3.0 N:Clark;Andrew G.; FN:Andrew G. Clark X-ORCID:https://orcid.org/0000-0001-7159-8511 END:VCARD BEGIN:VCARD VERSION:3.0 N:Xu;Xuejiao; FN:Xuejiao Xu END:VCARD BEGIN:VCARD VERSION:3.0 N:Langmüller;Anna M.; FN:Anna M. Langmüller X-ORCID:https://orcid.org/0000-0002-6102-8862 END:VCARD BEGIN:VCARD VERSION:3.0 N:Metzloff;Matthew; FN:Matthew Metzloff X-ORCID:https://orcid.org/0000-0002-6108-5031 END:VCARD https://w3id.org/kim/hcrt/scheme https://w3id.org/kim/hcrt/text Textdokument Text LOMv1.0 yes https://creativecommons.org/licenses/by/4.0 application/pdf 958088 https://phaidra.vetmeduni.ac.at/o:1864 https://phaidra.vetmeduni.ac.at/api/object/o:1864/thumbnail LOMv1.0 discipline