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RNA Innovation Seminar: Zhonggang Hou & Jun Hee Lee
Monday, November 8 @ 4:00 pm - 5:00 pm
“Harnessing diverse compact CRISPR-Cas3 for long-range genome engineering”
Zhonggang Hou, Ph.D.
ABSTRACT: Leading CRISPR-Cas technologies employ Cas9 and Cas12 enzymes that generate RNA-guided dsDNA breaks. Yet, the most abundant microbial adaptive immune systems, Type I CRISPR, is under-exploited for eukaryotic application. I will discuss our effort on adopting the first minimal CRISPR-Cas3 from Neisseria Type I-C system, to create targeted large chromosomal deletions in human cells. RNP delivery of the processive Cas3 nuclease and target recognition complex Cascade, gave up to 90% editing efficiency. Unexpectedly, Type I-C Cascade assembly in bacteria requires a previous unknown internal translation product Cas11 from within the cas8 gene. Our data show that expression of a separately encoded Cas11 is the key to enable plasmid- and mRNA- based editing in human cells. We demonstrate that “supplying cas11” is a universal strategy to harness divergent and streamlined Type I-C, I-D and I-B editors with distinct PAM preferences and guide orthogonality. Our findings expand the CRISPR toolbox for long-range genome engineering.
KEYWORDS: CRISPR; genome editing; Cascade; Cas3; Cas11; DNA targeting; crRNA; large deletion; Neisseria; genome engineering
“Microscopic Examination of Spatial Transcriptome through Seq-Scope”
Jun Hee Lee, Ph.D.
Molecular & Integrative Physiology
KEYWORDS: Spatial Transcriptomics, Seq-Scope