TY - JOUR
T1 - A multiplexed, confinable CRISPR/Cas9 gene drive can propagate in caged Aedes aegypti populations
AU - Anderson, Michelle A. E.
AU - Gonzalez, Estela
AU - Edgington, Matthew P.
AU - Ang, Joshua X. D.
AU - Purusothaman, Deepak Kumar
AU - Shackleford, Lewis
AU - Nevard, Katherine
AU - Verkuijl, Sebald A. N.
AU - Harvey-Samuel, Timothy
AU - Leftwich, Philip T.
AU - Esvelt, Kevin
AU - Alphey, Luke
N1 - Data availability statement: All raw reads from amplicon-Seq generated in this study were submitted to NCBI SRA with the accession number PRJNA741076. Addgene plasmid # 52891; http://n2t.net/addgene:52891; RRID:Addgene_52891. Plasmid sequences are available from NCBI OP728003 https://www.ncbi.nlm.nih.gov/nuccore/OP728003, OP728004, OP728005 https://www.ncbi.nlm.nih.gov/nuccore/OP728005. The remaining data generated for this study is available in the Supplementary dataset 1 file. Source data are provided with this paper.
Code availability: Details are given in the paper.
Funding information: M.A.E.A., E.G., J.X.D.A., L.S., K.N., S.A.N.V., and L.A. were funded through a Defense Advanced Research Projects Agency (DARPA) award [N66001-17-2-4054] to Kevin Esvelt at MIT. MPE and PTL were supported by the Wellcome Trust [110117/Z/15/Z]. L.A. and T.H.S. were funded by the UK Biotechnology and Biological Sciences Research Council [BBS/E/I/00007033, BBS/E/I/00007038, and BBS/E/I/00007039 to The Pirbright Institute]. DKP’s PhD studentship was funded by The Pirbright Institute. The views, opinions and/or findings expressed are those of the authors and should not be interpreted as representing the official views or policies of the U.S. Government. The funders had no role in study design, data collection and analysis, decision to publish, or preparation of the manuscript.
Rights retention statement: For the purpose of Open Access, the author has applied a CC BY public copyright license to any Author Accepted Manuscript (AAM) version arising from this submission.
PY - 2024/1/25
Y1 - 2024/1/25
N2 - Aedes aegypti is the main vector of several major pathogens including dengue, Zika and chikungunya viruses. Classical mosquito control strategies utilizing insecticides are threatened by rising resistance. This has stimulated interest in new genetic systems such as gene drivesHere, we test the regulatory sequences from the Ae. aegypti benign gonial cell neoplasm (bgcn) homolog to express Cas9 and a separate multiplexing sgRNA-expressing cassette inserted into the Ae. aegypti kynurenine 3-monooxygenase (kmo) gene. When combined, these two elements provide highly effective germline cutting at the kmo locus and act as a gene drive. Our target genetic element drives through a cage trial population such that carrier frequency of the element increases from 50% to up to 89% of the population despite significant fitness costs to kmo insertions. Deep sequencing suggests that the multiplexing design could mitigate resistance allele formation in our gene drive system.
AB - Aedes aegypti is the main vector of several major pathogens including dengue, Zika and chikungunya viruses. Classical mosquito control strategies utilizing insecticides are threatened by rising resistance. This has stimulated interest in new genetic systems such as gene drivesHere, we test the regulatory sequences from the Ae. aegypti benign gonial cell neoplasm (bgcn) homolog to express Cas9 and a separate multiplexing sgRNA-expressing cassette inserted into the Ae. aegypti kynurenine 3-monooxygenase (kmo) gene. When combined, these two elements provide highly effective germline cutting at the kmo locus and act as a gene drive. Our target genetic element drives through a cage trial population such that carrier frequency of the element increases from 50% to up to 89% of the population despite significant fitness costs to kmo insertions. Deep sequencing suggests that the multiplexing design could mitigate resistance allele formation in our gene drive system.
UR - http://www.scopus.com/inward/record.url?scp=85183000001&partnerID=8YFLogxK
U2 - 10.1038/s41467-024-44956-2
DO - 10.1038/s41467-024-44956-2
M3 - Article
C2 - 38272895
AN - SCOPUS:85183000001
VL - 15
JO - Nature Communications
JF - Nature Communications
SN - 2041-1723
M1 - 729
ER -
