Improved methods for the generation of human gene knockout and knockin cell lines.
Improved methods for the generation of human gene knockout and knockin cell lines.
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DOI：
10.1093/nar/gni160
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发表时间：
2005-10-07
影响因子：
14.9
通讯作者：
Bunz F
中科院分区：
文献类型：
--
作者：
Topaloglu O;Hurley PJ;Yildirim O;Civin CI;Bunz F
Recent studies have demonstrated the utility of recombinant adeno-associated viral (rAAV) vectors in the generation of human knockout cell lines. The efficiency with which such cell lines can be generated using rAAV, in comparison with more extensively described plasmid-based approaches, has not been directly tested. In this report, we demonstrate that targeting constructs delivered by rAAV vectors were nearly 25-fold more efficient than transfected plasmids that target the same exon. In addition, we describe a novel vector configuration which we term the synthetic exon promoter trap (SEPT). This targeting element further improved the efficiency of knockout generation and uniquely facilitated the generation of knockin alterations. An rAAV-based SEPT targeting construct was used to transfer a mutant CTNNB1 allele, encoding an oncogenic form of β-catenin, from one cell line to another. This versatile method was thus shown to facilitate the efficient integration of small, defined sequence alterations into the chromosomes of cultured human cells.
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影响因子：
46.9
作者：
Hirata, R;Chamberlain, J;Russell, DW
通讯作者：
Russell, DW
影响因子：
14.9
作者：
Kohli, M;Rago, C;Vogelstein, B
通讯作者：
Vogelstein, B
影响因子：
56.9
作者：
Bunz, F;Dutriaux, A;Vogelstein, B
通讯作者：
Vogelstein, B
影响因子：
5.3
作者：
HANSON, KD;SEDIVY, JM
通讯作者：
SEDIVY, JM
DOI：
10.1023/b:scam.0000007137.28557.73
发表时间：
1999-01-01
期刊：
SOMATIC CELL AND MOLECULAR GENETICS
影响因子：
--
作者：
Yáñez, RJ;Porter, ACG
通讯作者：
Porter, ACG