Crispr-cas component systems, methods and compositions for sequence manipulation

Abstract

The invention provides for systems, methods, and compositions for manipulation of sequences and/or activities of target sequences. Provided are vectors and vector systems, some of which encode one or more components of a CRISPR complex, as well as methods for the design and use of such vectors. Also provided are methods of directing CRISPR complex formation in eukaryotic cells and methods for selecting specific cells by introducing precise mutations utilizing the CRISPR/Cas system.

What is claimed is: 
     
         1 . A method comprising:
 introducing into, or expressing in, a eukaryotic cell having a DNA molecule,   (I) a Cas9 protein or a nucleotide sequence encoding the Cas9 protein, and   (II) RNA or a nucleotide sequence encoding the RNA, the RNA comprising:
 (a) a first RNA comprising a first ribonucleotide sequence and a second ribonucleotide sequence, and 
 (b) a second RNA, 
   wherein (II) (a) and (II) (b) are fused to one another or are covalently linked to one another with intervening nucleotides; and   wherein, in the eukaryotic cell, the second RNA forms an RNA duplex with the second ribonucleotide sequence, and the first ribonucleotide sequence directs the Cas9 protein to a target sequence of the DNA molecule, and the DNA molecule is cleaved or edited or at least one product of the DNA molecule is altered.