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Split versions of Cleave and Rescue selfish genetic elements for measured self limiting gene drive

Oberhofer, Georg and Ivy, Tobin and Hay, Bruce A. (2021) Split versions of Cleave and Rescue selfish genetic elements for measured self limiting gene drive. PLoS Genetics, 17 (2). Art. No. e1009385. ISSN 1553-7390. https://resolver.caltech.edu/CaltechAUTHORS:20200710-150639726

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Abstract

Gene drive elements promote the spread of linked traits, providing methods for changing the composition or fate of wild populations. Drive mechanisms that are self-limiting are attractive because they allow control over the duration and extent of trait spread in time and space, and are reversible through natural selection as drive wanes. Self-sustaining Cleave and Rescue (ClvR) elements include a DNA sequence-modifying enzyme such as Cas9/gRNAs that disrupts endogenous versions of an essential gene, a tightly linked recoded version of the essential gene resistant to cleavage (the Rescue), and a Cargo. ClvR spreads by creating loss-of-function (LOF) conditions in which those without ClvR die because they lack functional copies of the essential gene. We use modeling to show that when the Rescue-Cargo and one or both components required for LOF allele creation (Cas9 and gRNA) reside at different locations (split ClvR), drive of Rescue-Cargo is self-limiting due to a progressive decrease in Cas9 frequency, and thus opportunities for creation of LOF alleles, as spread occurs. Importantly, drive strength and duration can be extended in a measured manner—which is still self-limiting—by moving the two components close enough to each other that they experience some degree of linkage. With linkage, Cas9 transiently experiences drive by hitchhiking with Rescue-Cargo until linkage disequilibrium between the two disappears, a function of recombination frequency and number of generations, creating a novel point of control. We implement split ClvR in Drosophila, with key elements on different chromosomes. Cargo/Rescue/gRNAs spreads to high frequency in a Cas9-dependent manner, while the frequency of Cas9 decreases. These observations show that measured, transient drive, coupled with a loss of future drive potential, can be achieved using the simple toolkit that make up ClvR elements—Cas9 and gRNAs and a Rescue/Cargo.


Item Type:Article
Related URLs:
URLURL TypeDescription
https://doi.org/10.1371/journal.pgen.1009385DOIArticle
https://doi.org/10.1101/2020.07.09.196253DOIDiscussion Paper
ORCID:
AuthorORCID
Oberhofer, Georg0000-0003-0930-1996
Hay, Bruce A.0000-0002-5486-0482
Alternate Title:2-Locus ​Cleave and Rescue ​selfish elements harness a recombination rate-dependent generational clock for self limiting gene drive
Additional Information:© 2021 Oberhofer et al. This is an open access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited. Received: August 13, 2020; Accepted: January 28, 2021; Published: February 18, 2021. Stocks obtained from the Bloomington Drosophila Stock Center (NIH P40OD018537) were used in this study. Data Availability Statement: All relevant data are within the manuscript and its Supporting Information files. This work was carried out with support to B.A.H. from the US Department of Agriculture,National Institute of Food and Agriculture specialty crop initiative under USDA NIFA Award No. 2012-51181-20086, and Caltech. G.O. was supported by a Baxter Foundation Endowed Senior Postdoctoral Fellowship. T.I. was supported by NIH training grant 5T32GM007616-39. The funders had no role decision to publish, or preparation of the manuscript. Competing interests: The authors have filed patent applications on ClvR and related technologies (U.S. Application No. 15/970,728 and No. 16/673,823). Author Contributions: Conceptualization: Georg Oberhofer, Tobin Ivy, Bruce A. Hay. Funding acquisition: Georg Oberhofer, Bruce A. Hay. Investigation: Georg Oberhofer, Tobin Ivy, Bruce A. Hay. Methodology: Georg Oberhofer, Tobin Ivy. Software: Tobin Ivy. Validation: Georg Oberhofer, Tobin Ivy, Bruce A. Hay. Visualization: Georg Oberhofer, Tobin Ivy, Bruce A. Hay. Writing – original draft: Georg Oberhofer, Bruce A. Hay. Writing – review & editing: Georg Oberhofer, Tobin Ivy, Bruce A. Hay.
Funders:
Funding AgencyGrant Number
NIHP40OD018537
Department of Agriculture2012-51181-20086
CaltechUNSPECIFIED
Donald E. and Delia B. Baxter FoundationUNSPECIFIED
NIH Predoctoral Fellowship5T32GM007616-39
Issue or Number:2
Record Number:CaltechAUTHORS:20200710-150639726
Persistent URL:https://resolver.caltech.edu/CaltechAUTHORS:20200710-150639726
Official Citation:Oberhofer G, Ivy T, Hay BA (2021) Split versions of Cleave and Rescue selfish genetic elements for measured self limiting gene drive. PLoS Genet 17(2): e1009385. https://doi.org/ 10.1371/journal.pgen.1009385
Usage Policy:No commercial reproduction, distribution, display or performance rights in this work are provided.
ID Code:104337
Collection:CaltechAUTHORS
Deposited By: Tony Diaz
Deposited On:10 Jul 2020 22:28
Last Modified:12 Mar 2021 22:05

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