title: Intracellular delivery of artificial transcription factors fused to the protein transduction domain of HIV-1 Tat.
abstract: Protein transduction domains (PTDs), such as the TAT peptide derived from HIV Tat protein, may transduce macromolecules into cells. In the present study, the TAT peptide-fused artificial transcription factors (ATFs) were generated by fusion of the N-terminal TAT peptide with SV40 promoter-targeted three-fingered C2H2 zinc finger proteins and the KRAB transcriptional repression domain. The fusion proteins were then expressed in an E .coli system and purified by Ni-NTA affinity chromatography. The purified fusion proteins were tested on mammalian cell lines CHO DG44 and L929. TAT-ATF-S, which contains the zinc fingers that bind to the SV40 promoter with high specificity, exhibited the desired transcriptional repression activity to the reported genes, indicating the successful cellular delivery and desired conformation of TAT-ATF-S. Our study has provided a new strategy for intracellular ATF delivery.
authors: Beijing Institute of Biotechnology, 20 Dongdajie Street, Fengtai District, Beijing 100071, People's Republic of China. zhao.xinghui@gmail.com
authors: Xinghui
authors: X
authors: Zhao
authors: Yunzhu
authors: Y
authors: Dong
authors: Zhanzhong
authors: Z
authors: Zhao
authors: Junwei
authors: J
authors: Guo
authors: Jie
authors: J
authors: Liu
authors: Peitang
authors: P
authors: Huang
authors: Dayong
authors: D
authors: Dong
authors: Hongyan
authors: H
authors: Fan
authors: Qiang
authors: Q
authors: Guo
authors: Xiuxu
authors: X
authors: Yang
authors: Junjie
authors: J
authors: Xu
authors: Jianmin
authors: J
authors: Li
authors: Ling
authors: L
authors: Fu
authors: Wei
authors: W
authors: Chen
mesh_terms: D000818:Animals; D001665:Binding Sites; D016466:CHO Cells; D006224:Cricetinae; D003412:Cricetulus; D015696:Gene Products, tat / Q000235:genetics; D015497:HIV-1 / Q000235:genetics / Q000378:metabolism*; D017434:Protein Structure, Tertiary; D011993:Recombinant Fusion Proteins / Q000235:genetics* / Q000378:metabolism; D014157:Transcription Factors / Q000235:genetics / Q000378:metabolism; D016335:Zinc Fingers; D054322:tat Gene Products, Human Immunodeficiency Virus / Q000235:genetics* / Q000378:metabolism
chemical_list: D015696:Gene Products, tat; D011993:Recombinant Fusion Proteins; D014157:Transcription Factors; D054322:tat Gene Products, Human Immunodeficiency Virus