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

ABSTRACT: Engineered zinc-finger transcription 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 protocol, we describe a user-friendly ZF technology suitable for the creation of complex libraries and the construction of customized ZF-TFs. The new technology described here simplifies the building of ZF 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 9-11 d.

SUBMITTER: Gonzalez B

PROVIDER: S-EPMC2855653 | biostudies-other | 2010 Apr

REPOSITORIES: biostudies-other

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

Gonzalez Beatriz B Schwimmer Lauren J LJ Fuller Roberta P RP Ye Yongjun Y Asawapornmongkol Lily L Barbas Carlos F CF

Nature protocols 20100401 4

Engineered zinc-finger transcription 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 protocol, we describe a user-friendly ZF technology suitable for the creation of complex ...[more]

PMID: 20360772

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