MAY 08, 2019 6:00 AM PDT

Upgrading the CRISPR Toolbox - New Strategies Sharpen the CRISPR-Cas9 Scissors Utilizing Visualization and Chromatic Accessibility

Sponsored by: MilliporeSigma
C.E. Credits: P.A.C.E. CE Florida CE
Speakers
  • R&D Scientist, MilliporeSigma
    Biography
      Michelle has extensive R&D and product development experiences across different industries. She started her career at Bayer Crop Science (formerly Monsanto), developing new agricultural traits for insect control. After joining MilliporeSigma in 2015, her research focused on the function of cutting-edge genome editing tools. She led the research on two novel CRISPR technologies, "proxy-CRISPR" and "CRISPR-chrom," which have been published in Nature Communications and The Crispr Journal. Michelle earned her B.S. in Biotechnology from Nanjing Agricultural University and M.S. in Cellular and Molecular Biology from University of Missouri, St. Louis.
    • Senior R&D Scientist, MilliporeSigma
      Biography
        Dr. Yanfang Jiang obtained her PhD degree in microbiology from Department of Microbiology, University of Illinois at Urbana-Champaign, in the laboratory of Dr. John E. Cronan, who is a member of the National Academy of Sciences. Her thesis work focused on lipid and fatty metabolism and fatty acid derived cofactors in bacteria. She completed her postdoctoral training in Washington University in St. Louis, worked on glucose sensing and transport in yeast S. cerevisiae with Dr. Mark Johnston in the Department of Genetics and Center for Genome Sciences. She later worked in Dr. David Wang's laboratory in the Department of Molecular Microbiology in Washington University, involved in discovery and characterization of the first viral infection in nematodes. Dr. Jiang joined the Genome Engineering R&D Group in MilliporeSigma in October 2015 and has been primarily working on developing CRISPR-Cas9 protein products.

      Abstract

      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 


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