Precise genome editing with A.s. Cas12a (Cpf1) Ultra: a mutant protein with the reliability of S.p. Cas9
CRISPR-Cas12a (Cpf1) is an RNA-guided DNA endonuclease that recognizes TTTV (V = A/G/C) PAM sites, thereby permitting editing in organisms with AT-rich genomes. It has been widely reported that the wild-type A.s. Cas12a enzyme lacks the potency and site-to-site reliability of S.p. Cas9 when used to edit the human genome. Despite this shortcoming, A.s. Cas12a is an attractive option for genome editing due to its AT-rich PAM sequence, high fidelity, and its native reliance on a single short RNA guide. In this webinar, IDT staff scientist Dr Chris Vakulskas will talk about the development of a mutant protein, A.s. Cas12a (Cpf1) Ultra, which has dramatically improved cleavage activity that rivals S.p. Cas9 without altering its PAM specificity. The A.s. Cas12a (Cpf1) Ultra protein also demonstrates low-temperature cleavage activity superior to that of L.b. Cas12a, making it a universal Cas12a solution for genome editing in both animal and plant species. Finally, IDT research scientist Bernice Thommandru will describe optimal HDR methods and reagents for maximizing HDR rates when using either A.s. Cas12a (Cpf1) Ultra or S.p. Cas9.