HIGHLY COMPACT CAS9-BASED TRANSCRIPTIONAL REGULATORS FOR IN VIVO GENE REGULATION

摘要

Systems and methods for modulating gene expression are disclosed herein. The systems include a modified Cas9 protein having an HNH domain deletion, a guide RNA bound to the modified Cas9 protein, and at least one effector domain bound to the modified Cas9 protein, the guide RNA, or both. An AAV vector system for delivery of the gene expression modulation system includes a first expression cassette including a polynucleotide that encodes the modified Cas9 protein, a second expression cassette including a polynucleotide that encodes the guide RNA, and a third expression cassette including a polynucleotide that encodes the effector domain. The first, second, and third expression cassettes are located on one or more AAV vectors. Methods of modulating gene expression include introducing a system for modulating gene expression into a cell, hybridizing the guide RNA to a target gene, directing the effector domain to the target gene, and modulating transcription.