Patent ID: 8945839
Filing Date: 2015-02-03
Classification: A61K,C12N,C12Q

Abstract:
1. A method of altering expression of at least one gene product comprising introducing into a eukaryotic cell containing, and expressing a DNA molecule having a target sequence and encoding the gene product an engineered, non-naturally occurring Clustered Regularly Interspaced Short Palindromic Repeats (CRISPR)-CRISPR associated (Cas) system comprising one or more vectors comprising: a) a first regulatory element operable in a eukaryotic cell operably linked to at least one nucleotide sequence encoding a CRISPR-Cas system guide RNA that hybridizes with the target sequence; and b) a second regulatory element operable in a eukaryotic cell operably linked to a nucleotide sequence encoding a Type-II Cas9 protein, wherein: components (a) and (b) are located on same or different vectors of the system, the CRISPR-Cas system comprises one or more nuclear localization signals (NLSs), the guide RNA comprises a tracr sequence which is 30 or more nucleotides in length, and the Cas9 protein comprises one or more mutations in a catalytic domain, whereby the guide RNA targets and hybridizes to the target sequence, whereby expression of the at least one gene product is altered; and, wherein the Cas9 protein and the guide RNA do not naturally occur together.