RT Journal Article
SR Electronic
T1 Multiple loci of small effect confer wide variability in efficiency and resistance rate of CRISPR gene drive
JF bioRxiv
FD Cold Spring Harbor Laboratory
SP 447615
DO 10.1101/447615
A1 Jackson Champer
A1 Zhaoxin Wen
A1 Anisha Luthra
A1 Riona Reeves
A1 Joan Chung
A1 Chen Liu
A1 Yoo Lim Lee
A1 Jingxian Liu
A1 Emily Yang
A1 Philipp W. Messer
A1 Andrew G. Clark
YR 2018
UL http://biorxiv.org/content/early/2018/10/19/447615.abstract
AB Gene drives could allow for control of vector-borne diseases by directly suppressing vector populations or spreading genetic payloads designed to reduce pathogen transmission. CRISPR homing gene drives work by cleaving wild-type alleles, which are then converted to drive alleles by homology-directed repair, increasing the frequency of the drive in a population. However, resistance alleles can form when end-joining repair takes place in lieu of homology-directed repair. Such alleles cannot be converted to drive alleles, which would halt the spread of a drive through a population. To investigate the effects of natural genetic variation on resistance formation, we developed a CRISPR homing gene drive in Drosophila melanogaster and crossed it into the genetically diverse Drosophila Genetic Reference Panel (DGRP) lines, measuring several performance parameters. Most strikingly, resistance allele formation post-fertilization in the early embryo ranged from 7% to 79% among lines and averaged 42±18%. We performed a Genome-Wide Association Study (GWAS) using our results in the DGRP lines and found that the resistance and conversion rates were polygenic, with several genetic polymorphisms showing relatively weak association. RNAi knockdown of several of these genes confirmed their effect, but their small effect sizes implies that their manipulation will yield only modest improvements to the efficacy of gene drives.