Tracking genome engineering outcome at individual DNA breakpoints.
Tracking genome engineering outcome at individual DNA breakpoints.
复制标题
在单个DNA断点处跟踪基因组工程结果。
DOI：
10.1038/nmeth.1648
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发表时间：
2011-07-10
期刊：
影响因子：
48
通讯作者：
Scharenberg, Andrew M.
中科院分区：
文献类型：
--
作者：
Certo, Michael T.;Ryu, Byoung Y.;Annis, James E.;Garibov, Mikhail;Jarjour, Jordan;Rawlings, David J.;Scharenberg, Andrew M.
Site-specific genome engineering technologies are increasingly important tools in the post-genomic era, where biotechnological objectives often require organisms with precisely modified genomes. Rare-cutting endonucleases, through their capacity to create a targeted DNA strand break, are one of the most promising of these technologies. However, realizing the full potential of nuclease-induced genome engineering requires a detailed understanding of the variables that influence resolution of nuclease-induced DNA breaks. Here we present a genome engineering reporter system, designated Traffic Light, that supports rapid flow cytometric analysis of repair pathway choice at individual DNA breaks, quantitative tracking of nuclease expression and donor template delivery, and high throughput screens for factors that bias the engineering outcome. We applied the Traffic Light system to evaluate the efficiency and outcome of nuclease-induced genome engineering in human cell lines and identified strategies to facilitate isolation of cells in which a desired engineering outcome has occurred.
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DOI：
10.1126/science.1168683
发表时间：
2009-05-08
期刊：
Science (New York, N.Y.)
影响因子：
--
作者：
Shu X;Royant A;Lin MZ;Aguilera TA;Lev-Ram V;Steinbach PA;Tsien RY
通讯作者：
Tsien RY
影响因子：
20.3
作者：
Kustikova, OS;Wahlers, A;Fehse, B
通讯作者：
Fehse, B
影响因子：
10.5
作者：
Pierce, AJ;Hu, P;Jasin, M
通讯作者：
Jasin, M
影响因子：
14.9
作者：
Takeuchi, Ryo;Certo, Michael;Stoddard, Barry L.
通讯作者：
Stoddard, Barry L.
DOI：
10.1073/pnas.0810588106
发表时间：
2009-03-31
影响因子：
11.1
作者：
Smith, Audrey McConnell;Takeuchi, Ryo;Stoddard, Barry L.
通讯作者：
Stoddard, Barry L.