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Nucleic Acids Research Advance Access published online on July 21, 2008
Nucleic Acids Research, doi:10.1093/nar/gkn398
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© 2008 The Author(s)
This is an Open Access article distributed under the terms of the Creative Commons Attribution Non-Commercial License (http://creativecommons.org/licenses/by-nc/2.0/uk/) which permits unrestricted non-commercial use, distribution, and reproduction in any medium, provided the original work is properly cited.

Methods Online

Transduction of artificial transcriptional regulatory proteins into human cells

Chae-Ok Yun1, Hyun-Chul Shin2,3, Tae-Dong Kim4, Wan-Hee Yoon4, Yoon-A Kang1, Heung-Sun Kwon2, Seong Keun Kim2 and Jin-Soo Kim2,5,*

1Brain Korea 21 Project for Medical Sciences, Institute for Cancer Research, Yonsei Cancer Center, Yonsei University College of Medicine, 2ToolGen, Inc. Biotechnology Incubating Center, Gwanak-gu, Seoul, 3Department of Chemistry Education, Korea National University of Education, Chungbuk, 4College of Medicine, Chungnam National University, Daejon and 5Department of Chemistry, College of Natural Sciences, Seoul National University, Seoul, South Korea

*To whom correspondence should be addressed. Tel: 82-2-880-9327; Fax: 82-2-874-7455; Email: jskim01{at}snu.ac.kr

Received April 17, 2008. Revised June 5, 2008. Accepted June 5, 2008.

Protein transduction (PT) is a method for delivering proteins into mammalian cells. PT is accomplished by linking a small peptide tag—called a PT domain (PTD)—to a protein of interest, which generates a functional fusion protein that can penetrate efficiently into mammalian cells. In order to study the functions of a transcription factor (TF) of interest, expression plasmids that encode the TF often are transfected into mammalian cells. However, the efficiency of DNA transfection is highly variable among different cell types and is usually very low in primary cells, stem cells and tumor cells. Zinc-finger transcription factors (ZF-TFs) can be tailor-made to target almost any gene in the human genome. However, the extremely low efficiency of DNA transfection into cancer cells, both in vivo and in vitro, limits the utility of ZF-TFs. Here, we report on an artificial ZF-TF that has been fused to a well-characterized PTD from the human immunodeficiency virus-1 (HIV-1) transcriptional activator protein, Tat. This ZF-TF targeted the endogenous promoter of the human VEGF-A gene. The PTD-attached ZF-TF was delivered efficiently into human cells in vitro. In addition, the VEGF-A-specific transcriptional repressor retarded the growth rate of tumor cells in a mouse xenograft experiment.

The authors wish it to be known that, in their opinion, the first two authors should be regarded as joint First Authors.


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