University of Massachusetts Medical Center
University of Massachusetts Medical School
Clustered regularly-interspaced short palindromic repeats (CRISPR) systems, particularly Cas9, have become indispensable in the fields of biology as tools for the targeted modification of DNA. However, the new CRISPR enzyme Cpf1 provides a more robust platform, with the novel ability to process pre-CRISPR RNAs and generate staggered cuts in double-stranded DNA. Cpf1 has been used to introduce mutations to various ends, but it has only just begun to be used for gene regulation. To develop Cpf1 into a tool for such studies, the DNA and RNA catalytic domains in three Cpf1 species (As, Fn, Lb) were inactivated. The DNase-dead mutants were then fused to transcriptional effectors (VPR/KRAB), and the Lb-VPR construct was introduced into HEK293T cells to investigate its effect on CXCR4 expression.
Worcester Polytechnic Institute
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Chemistry and Biochemistry
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