A CRISPR homing gene drive targeting a haplolethal gene removes resistance alleles and successfully spreads through a cage population

Engineered gene drives are being explored as a new strategy in the fight against vector-borne diseases due to their potential for rapidly spreading genetic modifications through a population. However, CRISPR-based homing gene drives proposed for this purpose have faced a major obstacle in the format...

Full description

Saved in:
Bibliographic Details
Published in:Proceedings of the National Academy of Sciences - PNAS 2020-09, Vol.117 (39), p.24377-24383
Main Authors: Champer, Jackson, Yang, Emily, Lee, Esther, Liu, Jingxian, Clark, Andrew G., Messer, Philipp W.
Format: Article
Language:English
Subjects:
Alleles
Animals
Biological Sciences
Cages
CRISPR
CRISPR-Cas Systems
Drosophila melanogaster - genetics
Drosophila melanogaster - physiology
Drosophila Proteins - genetics
Female
Gene Editing
Genetic modification
Germ Cells - cytology
Heterozygotes
Homing
Infectious diseases
Male
Models, Genetic
Pedigree
Population genetics
Progeny
RNA, Guide, CRISPR-Cas Systems - genetics
Vector-borne diseases
Citations: Items that this one cites
Items that cite this one
Online Access:Get full text
Tags: Add Tag
No Tags, Be the first to tag this record!
Description
Summary:Engineered gene drives are being explored as a new strategy in the fight against vector-borne diseases due to their potential for rapidly spreading genetic modifications through a population. However, CRISPR-based homing gene drives proposed for this purpose have faced a major obstacle in the formation of resistance alleles that prevent Cas9 cleavage. Here, we present a homing drive in Drosophila melanogaster that reduces the prevalence of resistance alleles below detectable levels by targeting a haplolethal gene with two guide RNAs (gRNAs) while also providing a rescue allele. Resistance alleles that form by end-joining repair typically disrupt the haplolethal target gene and are thus removed from the population because individuals that carry them are nonviable. We demonstrate that our drive is highly efficient, with 91% of the progeny of drive heterozygotes inheriting the drive allele and with no functional resistance alleles observed in the remainder. In a large cage experiment, the drive allele successfully spread to all individuals within a few generations. These results show that a haplolethal homing drive can provide an effective tool for targeted genetic modification of entire populations.
ISSN:0027-8424
1091-6490
DOI:10.1073/pnas.2004373117