Precision Genome Editing in Sugarcane

Genome editing with a designer nuclease such as CRISPR/Cas9 enabling precise targeting and introduction of double stranded DNA breaks has been described in various plant genomes. Subsequent cellular repair mechanisms, predominantly non-homologous end joining (NHEJ), act as critical steps to endogenous gene editing or correction. However, there is very limited control over these mechanisms, which generate an abundance of random insertions and deletions (indels). Frameshift mutations associated with these indels of unspecified size and sequence might result in loss of function phenotypes of agronomic importance. Gain of function mutations, on the other hand, generally requires precise editing of the target locus with the aid of a homologous template and involves the cellular homology directed repair (HDR) mechanism. We will present an HDR mediated genome editing approach in sugarcane and will discuss the impact of alternative guide RNA’s and delivered amount of components.