CRISPR in cyanobacteria: A system for genomic editing in Synechocystis

Abstract

CRISPR-system genomic editing is perhaps one of the most profound and far-reaching technological breakthroughs of the last decade; however, we have still yet to surpass the earliest stages of understanding this new technology. Through DNA manipulation achieved by the use of restriction digestion, Gibson Assembly, E. coli transformation, triparental conjugation and we established a foundation from which CRISPR experimentation can be achieved cheaply and easily in our laboratory in the Department of Microbiology and Molecular Genetics at Oklahoma State University. This foundation can be used as a template for further research and has been tested through the genetic knockout of the cpcBACCD operon (Figure 1) encoding the major light-harvesting antenna system of the cyanobacterial model organism Synechocystis sp. PCC6803.