Francisella novicida Cas9 interrogates genomic DNA with very high specificity and can be used for mammalian genome editing

Acharya S, Mishra A, Paul D, et al.

Proceedings of the National Academy of Sciences of the United States of America 2019, vol: 116(42) doi: 10.1073/pnas.1818461116

Abstract

Genome editing using the CRISPR/Cas9 system has been used to make precise heritable changes in the DNA of organisms. Although the widely used Streptococcus pyogenes Cas9 (SpCas9) and its engineered variants have been efficiently harnessed for numerous gene-editing applications across different platforms, concerns remain regarding their putative off-targeting at multiple loci across the genome. Here we report that Francisella novicida Cas9 (FnCas9) shows a very high specificity of binding to its intended targets and negligible binding to off-target loci. The specificity is determined by its minimal binding affinity with DNA when mismatches to the target single-guide RNA (sgRNA) are present in the sgRNA:DNA heteroduplex. FnCas9 produces staggered cleavage, higher homology-directed repair rates, and very low nonspecific genome editing compared to SpCas9. We demonstrate FnCas9-mediated correction of the sickle cell mutation in patient-derived induced pluripotent stem cells and propose that it can be used for precise therapeutic genome editing for a wide variety of genetic disorders.

View Publication
 

Topics: Gene Therapy, Monolith – MicroScale Thermophoresis, MST,  Publications

 

 

 

Previous
What binding affinity and binding kinetics tell you about protein-ligand interactions
What binding affinity and binding kinetics tell you about protein-ligand interactions

Up next
Easily characterize the molecular mechanisms of neurodegenerative diseases
Easily characterize the molecular mechanisms of neurodegenerative diseases

Have a question about Monolith?

Contact Specialist
×

Sign up to receive
the latest NanoTemper news, product updates, event announcements and more

First Name
Last Name
Company Name
Country
State
Province
State
Region
State
Canton
Agree to Subscribe & Privacy Policy*
*I have fully read, understood and agree to the Privacy Policy. I accept the storing and processing of my personal data by NanoTemper as described in the Privacy Policy.

By completing this form, you provide us consent to contact you with educational content, news and information about our products, services and events. You may unsubscribe at any time.
Thank you!
Error - something went wrong!