Casilio: a versatile CRISPR-Cas9-Pumilio hybrid for gene regulation and genomic labeling OPEN

摘要

The CRISPR-Cas9 system has recently been widely adopted in genome editing due to its simplicity1,2,3. Nuclease-deficient mutant dCas9 protein can be fused to effector domains and the fusion proteins can be guided by sgRNAs to genomic sites to regulate gene expression or label chromosomes4,5,6,7,8,9,10. However, only one type of effector is applied in most experiments due to the exclusive sgRNA:Cas9 pairing. Moreover, multimerization by directly fusing multiple copies of effectors with dCas9 protein to achieve sufficient effector activity is technically challenging. RNA aptamer approaches utilizing viral RNA sequences such as MS2 and PP7 have been combined with the CRISPR-Cas9 system to provide tools with improved multiplexing and multimerization functionalities11,12.