CHENG

RNA-guided nuclease CRISPR/Cas9 is widely used in genome editing [1-5]. Nuclease-deficient mutant dCas9 can be repurposed as a versatile RNA-programmable DNA-binding domain that does not generate breaks on DNA [6]. CRISPR/dCas9 has been used to bind target DNA and block transcription, activate or repress genes, tether epigenetic factor such as histone acetyltransferase to modify epigenetic status [7-11] and visualize genomic loci in live cells by tethering fluorescent proteins to defined loci [12]. These CRISPR/dCas9-based enzymes (CRISPRzymes) enable researchers to perform functional studies in situ and in vivo without permanently changing genetic sequences... We have constructed an RNA-aptamer based approach called Casilio to improve both multiplexing and multimerization capabilities to CRISPRzyme toolkits [19]. Casilio is a hybrid system combining CRISPR/dCas 9 and Pum ilio RNA-binding domain (Fig 2). Pumilio/PUF RNA binding domain contains 8 sequence repeats each recognizing one..

Primary location: Tempe United States