COMPOSITIONS AND METHODS OF USE OF CRISPR-CAS SYSTEMS IN NUCLEOTIDE REPEAT DISORDERS

摘要

The invention provides for delivery, engineering and optimization of systems, methods, and compositions for manipulation of sequences and/or activities of target sequences especially for use as to nucleotide repeat disorders. Provided are delivery systems and tissues or organ which are targeted as sites for delivery especially for use as to nucleotide repeat disorders. Also provided are vectors and vector systems some of which encode one or more components of a CRISPR complex or system especially for use as to nucleotide repeat disorders, as well as methods for the design and of such. Also provided are methods of directing CRISPR complex or system formation in eukaryotic cells especially for use as to nucleotide repeat disorders including with consideration of specificity for target recognition and avoidance of toxicity and editing or modifying a target site in a genomic locus of interest to alter or improve the status of a disease or a condition.

主权项

1. A non-naturally occurring or engineered composition suitable for use in a cell having a defective nucleotide element or trinucleotide repeat or other nucleotide repeat element or nucleotide expansion, the composition comprising:
I. at least two CRISPR-Cas system RNA polynucleotide sequences operably linked to one or more first regulatory elements, wherein each polynucleotide sequence comprises:
(a) at least one first guide sequence capable of hybridizing to a target DNA sequence within the cell,(b) at least one tracr mate sequence, and(c) at least one tracr sequence, and wherein (a), (b) and (c) are arranged in a 5′ to 3′ orientation, II. a second regulatory element operably linked to a polynucleotide sequence encoding a CRISPR enzyme,
wherein parts I and II comprise a CRISPR-Cas system, and wherein said composition comprisesa CRISPR complex comprising the translated CRISPR enzyme complexed with (1) the guide sequence that is hybridized or is hybridizable to the target sequence, and (2) the tracr mate sequence that is hybridized to the tracr sequence,wherein the guide sequences directs sequence-specific binding of a CRISPR complex to the target DNA, and wherein each of the two or more guides sequences hybridize to target DNA sequences that together flank the element, repeat or expansion to thereby mediate excision of the element, repeat or expansion by the CRISPR complex.