MUTAGENESIS METHODS

摘要

In some embodiments, aspects of the disclosure provide methods and compositions that are useful for modifying (e.g., mutating) one or more alleles of a genomic locus within a cell. In some embodiments, methods and compositions described herein involve producing a chimeric spliced RNA molecule that includes a transcribed exon spliced to a nuclease interacting RNA segment. In some embodiments, the chimeric spliced RNA guides a DNA modifying enzyme (e.g., a nuclease) to a genomic locus in a cell resulting in modification of the locus.

主权项

1. A method of producing, in a eukaryotic cell, a target-specific RNA molecule capable of guiding a DNA nuclease to a genomic target, the method comprising introducing a recombinant nucleic acid into a eukaryotic cell, wherein the recombinant nucleic acid comprises a first nucleic acid region that encodes a splice acceptor site upstream of a second nucleic acid region that encodes an RNA segment capable of interacting with an RNA-guided DNA nuclease.