Compounds and methods for CRISPR / CAS-based genome editing by homologous recombination

摘要

The present invention relates to guide RNAs comprising adapter segments with one or more modifications, and their use in homologous recombination by the CRISPR / Cas system. The modified adapter segment is resistant to degradation by RNase H. The present invention also relates to a dual guide RNA strategy wherein the first guide RNA directs the Cas enzyme to perform a double-strand break in the first target sequence, and the second guide RNA directs the adapters attached to the donor polynucleotide And binds to a second target sequence that comprises fragments and is spaced from the first target sequence.