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The Casilio system can be used for gene regulation, epigenetic editing, and chromosomal labeling. It also offers a toolbox to investigate basic research in gene regulatory networks
About
The Casilio system consists of (1) dCas9 protein, (2) an sgRNA appended with one or more PUF-binding site(s) (sgRNA-PBS), and (3) an effector fused with a PUF domain (PUF fusion). The sgRNA-PBS specifies both DNA binding via its spacer sequence and effector recruitment via PBS. Casilio is currently listed in Addgene but only available to academic institutions. You can find out more here: https://www.addgene.org/crispr/albert-cheng/
Key Benefits
• Simultaneously regulates multiple target genes in a cell (activate one gene while repress another) • Modulates different gene expression levels via effector multimerization • Allows dual/multiple delivery of different effectors at a single target gene when multiple proteins are necessary to enhance the genetic/epigenetic modification (e.g., TET1 effector and DNA glycosylase NEIL2 to improve demethylation and activation of methylation-silenced genes)
Applications
The Casilio system can be used for the following: • Activation and repression of different endogenous gene expressions simultaneously in a cell • Enhancement of precise editing of DNA double stranded break caused by gene editing tools (CRISPR) by recruiting DNA repair proteins at the nuclease target site • Efficient epigenetic editing via dual/multi effector delivery • Live cell imaging of nonrepetitive genomic loci (e.g., track chromatin dynamics) with high spatiotemporal resolution