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Nature DOI:10.1038/nature17938

Continuous evolution of Bacillus thuringiensis toxins overcomes insect resistance.

Publication TypeJournal Article
Year of Publication2016
AuthorsBadran, AH, Guzov, VM, Huai, Q, Kemp, MM, Vishwanath, P, Kain, W, Nance, AM, Evdokimov, A, Moshiri, F, Turner, KH, Wang, P, Malvar, T, Liu, DR
Date Published2016 May 05
KeywordsAmino Acid Sequence, Animals, Bacillus thuringiensis, Bacterial Proteins, Bacteriophages, Biotechnology, Cadherins, Cell Death, Consensus Sequence, Crops, Agricultural, Directed Molecular Evolution, Endotoxins, Genetic Variation, Hemolysin Proteins, Insecticide Resistance, Insecticides, Molecular Sequence Data, Moths, Mutagenesis, Pest Control, Biological, Plants, Genetically Modified, Protein Binding, Protein Stability, Selection, Genetic

The Bacillus thuringiensis δ-endotoxins (Bt toxins) are widely used insecticidal proteins in engineered crops that provide agricultural, economic, and environmental benefits. The development of insect resistance to Bt toxins endangers their long-term effectiveness. Here we have developed a phage-assisted continuous evolution selection that rapidly evolves high-affinity protein-protein interactions, and applied this system to evolve variants of the Bt toxin Cry1Ac that bind a cadherin-like receptor from the insect pest Trichoplusia ni (TnCAD) that is not natively bound by wild-type Cry1Ac. The resulting evolved Cry1Ac variants bind TnCAD with high affinity (dissociation constant Kd = 11-41 nM), kill TnCAD-expressing insect cells that are not susceptible to wild-type Cry1Ac, and kill Cry1Ac-resistant T. ni insects up to 335-fold more potently than wild-type Cry1Ac. Our findings establish that the evolution of Bt toxins with novel insect cell receptor affinity can overcome insect Bt toxin resistance and confer lethality approaching that of the wild-type Bt toxin against non-resistant insects.


Alternate JournalNature
PubMed ID27120167
PubMed Central IDPMC4865400
Grant List / / Howard Hughes Medical Institute / United States
R01 EB022376 / EB / NIBIB NIH HHS / United States
R01EB022376 / EB / NIBIB NIH HHS / United States