CRISPR-BASED GENOME MODIFICATION AND REGULATION

摘要

The present invention provides RNA-guided endonucleases, which are engineered for expression in eukaryotic cells or embryos, and methods of using the RNA-guided endonuclease for targeted genome modification in eukaryotic cells or embryos. Also provided are fusion proteins, wherein each fusion protein comprises a CRISPR/Cas-like protein or fragment thereof and an effector domain. The effector domain can be a cleavage domain, an epigenetic modification domain, a transcriptional activation domain, or a transcriptional repressor domain. Also provided are methods for using the fusion proteins to modify a chromosomal sequence or regulate expression of a chromosomal sequence.

主权项

1. A method for decreasing expression of a chromosomal sequence in a cell or embryo, the method comprising introducing into the cell or embryo (a) at least one fusion protein or nucleic acid encoding at least one fusion protein, wherein the fusion protein comprises a Cas9 protein mutated in the RuvC and HNH domains so that it no longer possesses nuclease activity or a fragment thereof and a transcriptional repressor domain, and (b) at least one guide RNA or DNA encoding at least one guide RNA, wherein the at least one guide RNA guides the mutated Cas9 protein of the fusion protein to a target site in the chromosomal sequence and the transcriptional repressor domain of the fusion protein decreases expression of the chromosomal sequence.