Patent ID: 9738897
Date: 2017-08-22
CPC Classifications: C12N

Claim:
1. An in vitro method for assembling at least two nucleic acids, comprising: (a) contacting a first nucleic acid with a first nuclease agent, wherein the first nuclease agent comprises a Cas protein and a guide RNA (gRNA) (gRNA-Cas complex), a zinc finger nuclease, or a Transcription Activator-Like Effector Nuclease (TALEN), wherein the first nuclease agent cleaves the first nucleic acid at a first target site to produce a first digested nucleic acid with an overlapping end sequence shared by a second nucleic acid; (b) contacting the first digested nucleic acid and the second nucleic acid with an exonuclease to expose complementary sequences between the first digested nucleic acid and the second nucleic acid; and (c) assembling the two nucleic acid fragments generated from step (b), comprising: