	US 11,897,920 B2
	Tale RVD specifically recognizing DNA base modified by methylation and application thereof
Wensheng Wei, Beijing (CN); Chengqi Yi, Beijing (CN); Yuan Zhang, Beijing (CN); Shengjie Guo, Beijing (CN); Chenxu Zhu, Beijing (CN); and Lulu Liu, Beijing (CN)
Assigned to Peking University, Beijing (CN); and Edigene Inc., Beijing (CN)
Appl. No. 16/636,283
Filed by PEKING UNIVERSITY, Beijing (CN); and EDIGENE INC., Beijing (CN)
PCT Filed Aug. 4, 2017, PCT No. PCT/CN2017/095988 § 371(c)(1), (2) Date Feb. 3, 2020, PCT Pub. No. WO2019/024081, PCT Pub. Date Feb. 7, 2019.
Prior Publication US 2021/0054030 A1, Feb. 25, 2021
Int. Cl. C07K 14/195 (2006.01); C12N 9/22 (2006.01); C12N 15/113 (2010.01); C12N 15/66 (2006.01)

CPC C07K 14/195 (2013.01) [C12N 9/22 (2013.01); C12N 15/113 (2013.01); C12N 15/66 (2013.01); C07K 2319/00 (2013.01); C07K 2319/80 (2013.01); C12N 2310/20 (2017.05); C12N 2800/80 (2013.01)]

4 Claims

1. A method for detecting the presence of 5-methylcytosine (5mC) at a specific site of a target sequence in the genome of a cell, comprising:

(1) introducing a protein comprising transcription activator-like effectors (TALEs) into the cell, wherein the TALEs target the target sequence, and in the TALEs the repeat variable diresidue (RVD) identifying the specific site is HA;

(2) introducing a nuclease into the cell, wherein the targeted cleavage site of the nuclease is located in the TALEs target sequence; and

(3) detecting whether the target sequence is cleaved, thereby judging whether 5mC is present at the specific site of the target sequence; if the target sequence is not cleaved, then the TALEs bind to the target sequence, and the nuclease does not bind to the target sequence and cleave it, thereby 5mC is present at the specific site; if the target sequence is cleaved, then the TALEs do not bind to the target sequence, and the nuclease binds to the target sequence and cleaves it, thereby 5mC is not present at the specific site.