Human zinc fingers as building blocks in the construction of artificial transcription factors

• Published in: Nature biotechnology 2003-03, Vol.21 (3), p.275-280
• Main Authors: Kim, Jin-Soo, Bae, Kwang-Hee, Do Kwon, Young, Shin, Hyun-Chul, Hwang, Moon-Sun, Ryu, Eun-Hyun, Park, Kyung-Soon, Yang, Hyo-Young, Lee, Dong-ki, Lee, Yangsoon, Park, Jinwoo, Sun Kwon, Heung, Kim, Hyun-Won, Yeh, Byung-Il, Lee, Hyean-Woo, Hyung Sohn, Soon, Yoon, Joonho, Seol, Wongi
• Format: Article
• Language: English
• Subjects: Agriculture; Amino acids; Bioinformatics; Biological and medical sciences; Biomedical and Life Sciences; Biomedical Engineering/Biotechnology; Biomedicine; Biotechnology; Cloning; Deoxyribonucleic acid; DNA; DNA - chemistry; DNA - genetics; DNA - metabolism; DNA-Binding Proteins - biosynthesis; DNA-Binding Proteins - chemistry; DNA-Binding Proteins - genetics; Fundamental and applied biological sciences. Psychology; Gene Expression Regulation; Genes; Genetic engineering; Genetic technics; Genome, Human; Genomes; Humans; Life Sciences; Methods. Procedures. Technologies; Modification of gene expression level; Peptide Library; Plasmids; Prefabricated buildings; Protein Engineering - methods; Proteins; Recombinant Proteins - chemistry; Recombinant Proteins - genetics; Recombinant Proteins - metabolism; Transcription factors; Transcription Factors - biosynthesis; Transcription Factors - chemistry; Transcription Factors - genetics; Transcription, Genetic; Yeast; Yeasts - genetics; Yeasts - metabolism; Zinc; Zinc Fingers - genetics
• ISSN: 1087-0156; 1546-1696
• DOI: 10.1038/nbt796

We describe methods for generating artificial transcription factors capable of up- or downregulating the expression of genes whose promoter regions contain the target DNA sequences. To accomplish this, we screened zinc fingers derived from sequences in the human genome and isolated 56 zinc fingers with diverse DNA-binding specificities. We used these zinc fingers as modular building blocks in the construction of novel, sequence-specific DNA-binding proteins. Fusion of these zinc-finger proteins with either a transcriptional activation or repression domain yielded potent transcriptional activators or repressors, respectively. These results show that the human genome encodes zinc fingers with diverse DNA-binding specificities and that these domains can be used to design sequence-specific DNA-binding proteins and artificial transcription factors.