CRISPR-Cas9-mediated knockout of CYP79D1 and CYP79D2 in cassava attenuates toxic cyanogen production

Gomez, Michael A.; Berkoff, Kodiak C.; Gill, Baljeet K.; Iavarone, Anthony T.; Lieberman, Samantha E.; Ma, Jessica M.; Schultink, Alex; Karavolias, Nicholas G.; Wyman, Stacia K.; Chauhan, Raj Deepika; Taylor, Nigel J.; Staskawicz, Brian J.; Cho, Myeong-Je; Rokhsar, Daniel S.; Lyons, Jessica B.

doi:10.3389/fpls.2022.1079254

CRISPR-Cas9-mediated knockout of CYP79D1 and CYP79D2 in cassava attenuates toxic cyanogen production

Journal Article · Fri Mar 17 00:00:00 EDT 2023 · Frontiers in Plant Science

DOI:https://doi.org/10.3389/fpls.2022.1079254· OSTI ID:2470860

Gomez, Michael A. ^[1]; Berkoff, Kodiak C. ^[1]; Gill, Baljeet K. ^[1]; Iavarone, Anthony T. ^[1]; Lieberman, Samantha E. ^[1]; Ma, Jessica M. ^[1]; Schultink, Alex ^[2]; Karavolias, Nicholas G. ^[1]; Wyman, Stacia K. ^[1]; Chauhan, Raj Deepika ^[3]; Taylor, Nigel J. ^[4]; Staskawicz, Brian J. ^[1]; Cho, Myeong-Je ^[1]; Rokhsar, Daniel S. ^[5]; Lyons, Jessica B. ^[1]

University of California, Berkeley, CA (United States)
University of California, Berkeley, CA (United States); Fortiphyte, Berkeley, CA (United States)
Donald Danforth Plant Science Center, St. Louis, MO (United States); Pairwise, Durham, NC (United States)
Donald Danforth Plant Science Center, St. Louis, MO (United States)
University of California, Berkeley, CA (United States); Lawrence Berkeley National Laboratory (LBNL), Berkeley, CA (United States); USDOE Joint Genome Institute (JGI), Berkeley, CA (United States); Okinawa Institute of Science and Technology Graduate University, Onna (Japan); Chan Zuckerberg Biohub, San Francisco, CA (United States)

Cassava (Manihot esculenta) is a starchy root crop that supports over a billion people in tropical and subtropical regions of the world. This staple, however, produces the neurotoxin cyanide and requires processing for safe consumption. Excessive consumption of insufficiently processed cassava, in combination with protein-poor diets, can have neurodegenerative impacts. This problem is further exacerbated by drought conditions which increase this toxin in the plant. To reduce cyanide levels in cassava, we used CRISPR-mediated mutagenesis to disrupt the cytochrome P450 genes CYP79D1 and CYP79D2 whose protein products catalyze the first step in cyanogenic glucoside biosynthesis. Knockout of both genes eliminated cyanide in leaves and storage roots of cassava accession 60444; the West African, farmer-preferred cultivar TME 419; and the improved variety TMS 91/02324. Although knockout of CYP79D2 alone resulted in significant reduction of cyanide, mutagenesis of CYP79D1 did not, indicating these paralogs have diverged in their function. The congruence of results across accessions indicates that our approach could readily be extended to other preferred or improved cultivars. This work demonstrates cassava genome editing for enhanced food safety and reduced processing burden, against the backdrop of a changing climate.

View Accepted Manuscript (DOE)

Research Organization:: Lawrence Berkeley National Laboratory (LBNL), Berkeley, CA (United States)

Sponsoring Organization:: National Institutes of Health (NIH); USDOE

Grant/Contract Number:: AC02-05CH11231

OSTI ID:: 2470860

Journal Information:: Frontiers in Plant Science, Journal Name: Frontiers in Plant Science Vol. 13; ISSN 1664-462X

Publisher:: Frontiers Research FoundationCopyright Statement

Country of Publication:: United States

Language:: English

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59 BASIC BIOLOGICAL SCIENCES
CRISPR-Cas9
CRISPR-associated protein 9
CYP79D
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Manihot esculenta Crantz
Plant Sciences
cassava
climate resilience
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CRISPR-Cas9-mediated knockout of CYP79D1 and CYP79D2 in cassava attenuates toxic cyanogen production

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