发明名称 |
Genome editing using targeting endonucleases and single-stranded nucleic acids |
摘要 |
The present invention provides methods and kits for editing specific chromosomal sequences in cells. In particular, targeting endonucleases and single-stranded nucleic acids are used to edit the chromosomal sequence. |
申请公布号 |
US9512444(B2) |
申请公布日期 |
2016.12.06 |
申请号 |
US201113811884 |
申请日期 |
2011.07.22 |
申请人 |
Sigma-Aldrich Co. LLC |
发明人 |
Chen Fuqiang;Pruett-Miller Shondra M.;Davis Gregory D. |
分类号 |
C12N15/90;C12N15/85;C12N9/22 |
主分类号 |
C12N15/90 |
代理机构 |
Polsinelli PC |
代理人 |
Polsinelli PC |
主权项 |
1. A method for integrating at least one exogenous sequence into at least one chromosomal sequence in a cell, the method comprising:
a) introducing into the cell (i) at least one targeting endonuclease or nucleic acid encoding a targeting endonuclease, the targeting endonuclease being able to introduce a double-stranded break at a targeted cleavage site in the chromosomal sequence, (ii) at least one first single-stranded nucleic acid comprising a first region having substantial sequence identity to one side of the targeted cleavage site, (iii) at least one second single-stranded nucleic acid comprising a first region having substantial sequence identity to the other side of the targeted cleavage site, and (iv) at least one donor polynucleotide comprising the exogenous sequence that is flanked by a first sequence having substantial sequence identity to a second region of the first single-stranded nucleic acid and a second sequence having substantial sequence identity to a second region of the second single-stranded nucleic acid; and b) maintaining the cell under conditions such that exogenous sequence is integrated into the chromosomal sequence during repair of the double-stranded break introduced by the targeting endonuclease. |
地址 |
St. Louis MO US |