title: 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.
authors.affiliation: The Skaggs Institute for Chemical Biology, La Jolla, California, USA.
authors.forename: Beatriz
authors.initials: B
authors.lastname: Gonzalez
authors.forename: Lauren J
authors.initials: LJ
authors.lastname: Schwimmer
authors.forename: Roberta P
authors.initials: RP
authors.lastname: Fuller
authors.forename: Yongjun
authors.initials: Y
authors.lastname: Ye
authors.forename: Lily
authors.initials: L
authors.lastname: Asawapornmongkol
authors.forename: Carlos F
authors.initials: CF
authors.lastname: Barbas
mesh_terms: D000818:Animals; D001483:Base Sequence; D003001:Cloning, Molecular; D004926:Escherichia coli / Q000235:genetics; D005822:Genetic Vectors; D006801:Humans; D019151:Peptide Library; D010957:Plasmids / Q000235:genetics; D015202:Protein Engineering / Q000379:methods*; D011994:Recombinant Proteins / Q000235:genetics; D014157:Transcription Factors / Q000235:genetics; D016335:Zinc Fingers / Q000235:genetics*
chemical_list: D019151:Peptide Library; D011994:Recombinant Proteins; D014157:Transcription Factors