Journal
NATURE PROTOCOLS
Volume 5, Issue 4, Pages 791-810Publisher
NATURE PUBLISHING GROUP
DOI: 10.1038/nprot.2010.34
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Funding
- American Cancer Society Illinois Division
- US National Institutes of Health
- NATIONAL CANCER INSTITUTE [R21CA126664] Funding Source: NIH RePORTER
- NATIONAL INSTITUTE OF GENERAL MEDICAL SCIENCES [R01GM065059] Funding Source: NIH RePORTER
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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.
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