Genome editing with the CRISPR-Cas9 and CRISPR-Cas12a (Cpf1) systems has been a groundbreaking development in basic biomedical research, with significant implications for personalized medicine and the treatment of human diseases.
Investigators working with CRISPR-Cas9 and CRISPR-Cas12a (Cpf1) face 3 key challenges inherent to both systems: (1) efficient, low-toxicity delivery of CRISPR reagents; (2) target site specificity, with the potentially deleterious consequences of off-target editing; and (3) potent on-target editing, even at difficult target sites. Obtaining precise editing results begins with better tools and strategies for overcoming these inherent challenges.
In this webinar, Dr Christopher Vakulskas (IDT) discusses the solutions IDT scientists and their collaborators have developed to meet each key challenge. First, Dr Vakulskas presents data showing that delivery methods and guide RNA (gRNA) characteristics can significantly impact the success of genome editing experiments. Although plasmid-based in vitro transcription of CRISPR reagents may seem straightforward, recent findings show that these reagents can stimulate the cellular immune response. However, data obtained by Dr Vakulskas and his colleagues demonstrate that ribonucleoprotein (RNP) delivery [transfection of recombinant Cas9 enzyme complexed with a synthetic two-part gRNA system (crRNA and tracrRNA)] avoids unwanted stimulation of the cellular immune response. Dr Vakulskas then discusses the newest high-fidelity Cas9 enzymes, Alt-R HiFi S.p. Cas9 Nuclease V3 and Alt-R A.s. Cas12a (Cpf1) Nuclease V3 developed by IDT scientists. He describes the dual selection, rational design strategy used to generate this enzyme, and presents data showing that the Alt-R HiFi S.p. Cas9 Nuclease V3 dramatically reduces off-target editing, while maintaining strong on-target performance. Additional data regarding the Alt-R HiFi S.p. Cas9 Nuclease V3 is available in a recent Nature Medicine publication.
To learn more about our Alt-R CRISPR Systems, including the Alt-R HiFi S.p. Cas9 Nuclease V3, Alt-R A.s. Cas12a (Cpf1) Nuclease V3, and other CRISPR products, or to place custom orders, contact our scientific applications support group.