CRISPR-CAS COMPONENT SYSTEMS, METHODS AND COMPOSITIONS FOR SEQUENCE MANIPULATION
First Claim
1. An engineered CRISPR-Cas9 complex comprising a tracr mate sequence and a tracr sequence,wherein the tracr mate and tracr sequences have at least 50% sequence complementarity along the length of the shorter of the two when aligned, wherein the complementary portions of the tracr and tracr mate sequences hybridize and have a length comprising about 16 to about 28 nucleotides, and the complex is designed to target a DNA sequence.
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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.
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Citations
20 Claims
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1. An engineered CRISPR-Cas9 complex comprising a tracr mate sequence and a tracr sequence,
wherein the tracr mate and tracr sequences have at least 50% sequence complementarity along the length of the shorter of the two when aligned, wherein the complementary portions of the tracr and tracr mate sequences hybridize and have a length comprising about 16 to about 28 nucleotides, and the complex is designed to target a DNA sequence.
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11. A method of modifying a target DNA molecule, the method comprising:
- contacting a target DNA molecule having a target sequence with a complex comprising;
(a) a Cas9 protein; and (b) a DNA-targeting RNA comprising; (i) a targeter-RNA that hybridizes with the target sequence, and (ii) an activator-RNA that hybridizes with the targeter-RNA to form a double-stranded RNA (dsRNA) duplex of a protein-binding segment, wherein the activator-RNA hybridizes with the targeter-RNA to form a total of 10 to 15 base pairs, wherein said contacting takes place outside of a bacterial cell and outside of an archaeal cell, thereby resulting in modification of the target DNA molecule. - View Dependent Claims (12, 13, 14, 15, 16, 17, 18, 19, 20)
- contacting a target DNA molecule having a target sequence with a complex comprising;
Specification