Genetic conversion of a split-drive into a full-drive element

Gerard Terradas, Jared B. Bennett, Zhiqian Li, John M. Marshall and Ethan Bier ()
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Gerard Terradas: University of California, San Diego
Jared B. Bennett: University of California
Zhiqian Li: University of California, San Diego
John M. Marshall: University of California
Ethan Bier: University of California, San Diego

Nature Communications, 2023, vol. 14, issue 1, 1-10

Abstract: Abstract The core components of CRISPR-based gene drives, Cas9 and guide RNA (gRNA), either can be linked within a self-contained single cassette (full gene-drive, fGD) or be provided in two separate elements (split gene-drive, sGD), the latter offering greater control options. We previously engineered split systems that could be converted genetically into autonomous full drives. Here, we examine such dual systems inserted at the spo11 locus that are recoded to restore gene function and thus organismic fertility. Despite minimal differences in transmission efficiency of the sGD or fGD drive elements in single generation crosses, the reconstituted spo11 fGD cassette surprisingly exhibits slower initial drive kinetics than the unlinked sGD element in multigenerational cage studies, but then eventually catches up to achieve a similar level of final introduction. These unexpected kinetic behaviors most likely reflect differing transient fitness costs associated with individuals co-inheriting Cas9 and gRNA transgenes during the drive process.

Date: 2023
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DOI: 10.1038/s41467-022-35044-4

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