Modular system for the construction of zinc-finger libraries and proteins.

Modular system for the construction of zinc-finger libraries and proteins.
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DOI:
10.1038/nprot.2010.34
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发表时间:
2010-04
期刊:
影响因子:
14.8
通讯作者:
Barbas, Carlos F., III
Barbas, Carlos F., III
中科院分区:
生物学1区
文献类型:
--
作者:
Gonzalez, Beatriz;Schwimmer, Lauren J.;Fuller, Roberta P.;Ye, Yongjun;Asawapornmongkol, Lily;Barbas, Carlos F., III

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Engineered zinc-finger transcriptional factors (ZF-TF) are powerful tools to modulate the expression of specific genes. Complex libraries of ZF-TF can be delivered into cells to scan the genome for genes responsible for a particular phenotype or to select the most effective ZF-TF to regulate an individual gene. In both cases, the construction of highly representative and unbiased libraries is critical. In this work we describe a novel, user-friendly, zinc-finger technology suitable for the creation of complex libraries and the construction of customized ZF-TFs. The novel technology described here simplifies the building of zinc-finger libraries, avoids PCR-introduced bias, and ensures equal representation of every module. We also describe the construction of a customized ZF-TF that can be transferred to a number of expression vectors. This protocol can be completed in 9 to 11 days.
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