In this webinar, we will discuss our most recent additions to our CRISPR protein portfolio, the GFP-SpCas9 and GFP-eCas9 fusion proteins. The two GFP-Cas9 fusion proteins offer great visualization of intracellular transfected Cas9 proteins, which can be used for enrichment of hard-to-transfected cell lines. Our fusion proteins provide superior editing efficiency comparable to their non-fusion counterparts, outperforming similar products on the market with 10x fold more efficiency. 

Despite all the exciting promises it holds for advancing science and curing diseases, some hurdles remain before the full potential of this novel technology is unlocked, such as unwanted off target cleavage and low efficiencies at refractory sites.

The new CRISPR-chrom strategy developed by scientists at MilliporeSigma provides a quick and easy solution to improve genome modification efficiencies. This strategy works by fusing chromatin-modulating peptides (CMPs) to the Cas9 endonuclease. These peptides interact with chromatin and open the chromosome complexes for Cas9 to bind and cut. It was demonstrated that the CMP-fusion strategy improves the cleavage activities of Cas9 by several fold, particularly on previously identified difficult-to-cut loci.

In addition to the most widely adopted Streptococcus pyogenes Cas9 (SpCas9), this new strategy was also proven to be effective on various novel Cas9 orthologs, which hold the potential for better precision and new functionalities in genome editing research.

Learning Objectives: 

1. Cas9 proteins tagged with fluorescent markers allow easy visualization of Ribonuclearprotein (RNP) without sacrificing activity 
2. Formally inaccessible regions of the genome can now be targeted with proxy-CRISPR and CRISPR-chrom technologies