@article{5c9a43921d6f413bbc1569837871a502,
title = "Knockout of ABC transporter gene ABCA2 confers resistance to Bt toxin Cry2Ab in Helicoverpa zea",
abstract = "Evolution of pest resistance reduces the benefits of widely cultivated genetically engineered crops that produce insecticidal proteins derived from Bacillus thuringiensis (Bt). Better understanding of the genetic basis of pest resistance to Bt crops is needed to monitor, manage, and counter resistance. Previous work shows that in several lepidopterans, resistance to Bt toxin Cry2Ab is associated with mutations in the gene encoding the ATP-binding cassette protein ABCA2. The results here show that mutations introduced by CRISPR/Cas9 gene editing in the Helicoverpa zea (corn earworm or bollworm) gene encoding ABCA2 (HzABCA2) can cause resistance to Cry2Ab. Disruptive mutations in HzABCA2 facilitated the creation of two Cry2Ab-resistant strains. A multiple concentration bioassay with one of these strains revealed it had > 200-fold resistance to Cry2Ab relative to its parental susceptible strain. All Cry2Ab-resistant individuals tested had disruptive mutations in HzABCA2. We identified five disruptive mutations in HzABCA2 gDNA. The most common mutation was a 4-bp deletion in the expected Cas9 guide RNA target site. The results here indicate that HzABCA2 is a leading candidate for monitoring Cry2Ab resistance in field populations of H. zea.",
author = "Fabrick, {Jeffrey A.} and Heu, {Chan C.} and LeRoy, {Dannialle M.} and DeGain, {Ben A.} and Yelich, {Alex J.} and Unnithan, {Gopalan C.} and Yidong Wu and Xianchun Li and Yves Carri{\`e}re and Tabashnik, {Bruce E.}",
note = "Funding Information: This article was funded by USDA National Institute of Food and Agriculture, Agriculture and Food Research Initiative (2018-67013-27821). Funding Information: We thank Corteva Agriscience for providing Cry2A.127 used in this study. C.H. was supported, in part, by an appointment to the Agricultural Research Service (ARS) Research Participation Program admistered by the Oak Ridge Institute and Science Education (ORISE) through an interagency agreement between the U.S. Department of Energy (DOE) and the U.S. Department of Agriculture (USDA). ORISE is managed by Oak Ridge Associated Universities (ORAU) under DOE contract #DE-SC0014664. All opinions expressed in this paper are the authors' and do not necessarily reflect the policies and views of USDA, DOE, or ORAU/ORISE. Mention of trade names or commercial products in this article is solely for the purpose of providing specific information and does not imply recommendation or endorsement by the U.S. Department of Agriculture. USDA is an equal opportunity provider and employer. Funding Information: We thank Corteva Agriscience for providing Cry2A.127 used in this study. C.H. was supported, in part, by an appointment to the Agricultural Research Service (ARS) Research Participation Program admistered by the Oak Ridge Institute and Science Education (ORISE) through an interagency agreement between the U.S. Department of Energy (DOE) and the U.S. Department of Agriculture (USDA). ORISE is managed by Oak Ridge Associated Universities (ORAU) under DOE contract #DE-SC0014664. All opinions expressed in this paper are the authors' and do not necessarily reflect the policies and views of USDA, DOE, or ORAU/ORISE. Mention of trade names or commercial products in this article is solely for the purpose of providing specific information and does not imply recommendation or endorsement by the U.S. Department of Agriculture. USDA is an equal opportunity provider and employer. Funding Information: This is a cooperative investigation between USDA ARS and the University of Arizona, with J.A.F., Y.C., and B.E.T. receiving funding from USDA National Institute of Food and Agriculture (Agriculture and Food Research Initiative Program Grant #2018-67013-27821) to support this work. C.C.H., D.M.L., B.A.D., A.J.Y., G.C.U, and Y.W., declare no potential conflict of interest. J.A.F. is coauthor of a patent “Cadherin Receptor Peptide for Potentiating Bt Biopesticides” (patent numbers: US20090175974A1, US8354371, WO2009067487A2, WO2009067487A3). B.E.T. is a coauthor of a patent on modified Bt toxins, {"}Suppression of Resistance in Insects to Bacillus thuringiensis Cry Toxins, Using Toxins that do not Require the Cadherin Receptor{"} (patent numbers: CA2690188A1, CN101730712A, EP2184293A2, EP2184293A4, EP2184293B1, WO2008150150A2, WO2008150150A3). Bayer Crop Science, BASF, Corteva Agriscience, and Syngenta did not provide funding to support this work but may be affected financially by publication of this paper and have funded other work by J.A.F., Y.C., X.L., and/or B.E.T. Publisher Copyright: {\textcopyright} 2022, This is a U.S. Government work and not under copyright protection in the US; foreign copyright protection may apply.",
year = "2022",
month = dec,
doi = "10.1038/s41598-022-21061-2",
language = "English (US)",
volume = "12",
journal = "Scientific Reports",
issn = "2045-2322",
publisher = "Nature Publishing Group",
number = "1",
}