LAUSR

The Clustered Regularly Interspaced Short Palindromic Repeats (CRISPR)/Cas-mediated gene editing is a rapidly developing technique to investigate fundamental questions in Plant Science research and genetic improvement of plants. This system has been shown to work in over 70 different crop species from apple to wheat. Generation of site-speci fi c mutations in genes of interest requires expression of Cas (CRISPR-associated) nuclease and guide RNA (gRNA) in plant cells. Ef fi ciency of the CRISPR/Cas approach depends on several factors including but not limited to: (i) the type of Cas nuclease optimized to plant codons (ii) the presence and number of the NLS that can affect the Cas9 delivery to the cell nuclei, (iii) source and nature of promoters used to drive the expression of Cas genes, and (iv) an appropriate choice of target site and construction of gRNAs which can increase the mutation frequency. Finally, the editing success is contingent on the type of transgene delivery to the host/plant genome.