CRISPR/CAS9 SYSTEMS FOR PLANT GENOME EDITING
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Date
2016-04-28
Authors
Paul, Joseph W. III
Journal Title
Journal ISSN
Volume Title
Publisher
East Carolina University
Abstract
The increasing burden of the world population on agriculture requires the development of
more robust crops. Dissecting the basic biology that underlies plant susceptibility to pathogens
and drives crop loss will inform the design of better crops. One powerful tool for studying plants
at the molecular level is the RNA-programmed genome editing system comprised of a clustered
regularly-interspaced short palindromic repeats (CRISPR) encoded guide RNA (gRNA) and the
nuclease Cas9. CRISPR/Cas9 evolved as form of acquired immunity in bacteria and archaea but
can be re-engineered to edit and regulate eukaryotic genomes. The Cas9 nuclease encoded by
Streptococcus pyogenes (SpCas9) is most commonly used for genome editing but is
accompanied by strict requirements for nuclease activity. Here, we constructed and applied Cas9
systems for genome editing in plants with Cas9 derived from Streptococcus thermophiles
(StCas9) and Neisseria meningitidis (NmCas9) to expand on methods for gneome editing with
CRISPR/Cas9 in tobacco (Nicotiana benthamiana),. These results demonstrate an orthogonal
CRISPR/Cas9 system that could be further utilized for genome editing and simultaneous gene
regulation in plants, also providing a framework for synthetic transcriptional programming.
Translating this toolkit into crop species will provide increased control over plant genomes,
catalyzing new basic and applied discoveries to advance plant biology.