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Cleave and Rescue, a novel selfish genetic element and general strategy for gene drive

Oberhofer, Georg and Ivy, Tobin and Hay, Bruce A. (2019) Cleave and Rescue, a novel selfish genetic element and general strategy for gene drive. Proceedings of the National Academy of Sciences of the United States of America, 116 (13). pp. 6250-6259. ISSN 0027-8424. PMCID PMC6442612. http://resolver.caltech.edu/CaltechAUTHORS:20190214-105811281

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Abstract

There is great interest in being able to spread beneficial traits throughout wild populations in ways that are self-sustaining. Here, we describe a chromosomal selfish genetic element, CleaveR [Cleave and Rescue (ClvR)], able to achieve this goal. ClvR comprises two linked chromosomal components. One, germline-expressed Cas9 and guide RNAs (gRNAs)—the Cleaver—cleaves and thereby disrupts endogenous copies of a gene whose product is essential. The other, a recoded version of the essential gene resistant to cleavage and gene conversion with cleaved copies—the Rescue—provides essential gene function. ClvRenhances its transmission, and that of linked genes, by creating conditions in which progeny lacking ClvR die because they have no functional copies of the essential gene. In contrast, those who inherit ClvR survive, resulting in an increase in ClvR frequency. ClvR is predicted to spread to fixation under diverse conditions. To test these predictions, we generated a ClvR element in Drosophila melanogaster. ClvR^(tko) is located on chromosome 3 and uses Cas9 and four gRNAs to disrupt melanogaster technical knockout (tko), an X-linked essential gene. Rescue activity is provided by tko from Drosophila virilis. ClvR^(tko) results in germline and maternal carryover-dependent inactivation of melanogaster tko(>99% per generation); lethality caused by this loss is rescued by the virilis transgene; ClvR^(tko) activities are robust to genetic diversity in strains from five continents; and uncleavable but functional melanogaster tko alleles were not observed. Finally, ClvR^(tko) spreads to transgene fixation. The simplicity of ClvR suggests it may be useful for altering populations in diverse species.


Item Type:Article
Related URLs:
URLURL TypeDescription
https://doi.org/10.1073/pnas.1816928116DOIArticle
https://www.pnas.org/lookup/suppl/doi:10.1073/pnas.1816928116/-/DCSupplementalPublisherSupporting Information
https://www.ncbi.nlm.nih.gov/pmc/articles/PMC6442612PubMed CentralArticle
Additional Information:© 2019 the Author(s). Published by PNAS. This open access article is distributed under Creative Commons Attribution-NonCommercial-NoDerivatives License 4.0 (CC BY-NC-ND). Edited by James J. Bull, The University of Texas at Austin, Austin, TX, and approved January 7, 2019 (received for review October 2, 2018). PNAS published ahead of print February 13, 2019. Data Availability: All data are available in the main text or SI Appendix. ClvR^(tko) flies are available on request to labs that will meet or exceed containment guidelines outlined in ref. 61. We thank Marlene Biller and Alexander Sampson for technical assistance, and Jackson Champer and Andrew G. Clark for providing the GDL Drosophila strains. Stocks obtained from the Bloomington Drosophila Stock Center (NIH Grant P40OD018537) were used in this study. This work was carried out with support to B.A.H. from the US Department of Agriculture (USDA) National Institute of Food and Agriculture (NIFA) Specialty Crop Initiative, under USDA NIFA Award 2012-51181-20086. G.O. was supported by a research fellowship from the Deutsche Forschungsgemeinschaft (OB428/1-1). T.I. was supported by NIH Training Grant 5T32GM007616-39. Author contributions: G.O., T.I., and B.A.H. conceptualized the study; G.O., T.I., and B.A.H. provided the methodology; G.O. and B.A.H. investigated the study; T.I. provided mathematical modeling; G.O. and B.A.H. wrote the manuscript; G.O. and B.A.H. acquired funding; and G.O., T.I., and B.A.H. wrote the paper. Conflict of interest statement: The authors have filed patent applications on ClvR and related technologies (US Application No. 15/970,728). This article is a PNAS Direct Submission. This article contains supporting information online at www.pnas.org/lookup/suppl/doi:10.1073/pnas.1816928116/-/DCSupplemental.
Funders:
Funding AgencyGrant Number
NIHP40OD018537
Department of Agriculture (USDA)2012-51181-20086
Deutsche Forschungsgemeinschaft (DFG)OB428/1-1
NIH Predoctoral Fellowship5T32GM007616-39
Subject Keywords:gene drive; Cas9; population replacement; selfish genetic element
PubMed Central ID:PMC6442612
Record Number:CaltechAUTHORS:20190214-105811281
Persistent URL:http://resolver.caltech.edu/CaltechAUTHORS:20190214-105811281
Official Citation:Cleave and Rescue, a novel selfish genetic element and general strategy for gene drive. Georg Oberhofer, Tobin Ivy, Bruce A. Hay. Proceedings of the National Academy of Sciences. Mar 2019, 116 (13) 6250-6259; DOI: 10.1073/pnas.1816928116
Usage Policy:No commercial reproduction, distribution, display or performance rights in this work are provided.
ID Code:92935
Collection:CaltechAUTHORS
Deposited By: Tony Diaz
Deposited On:14 Feb 2019 19:37
Last Modified:09 Apr 2019 14:54

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