Human zinc fingers as building blocks in the construction of artificial transcription factors.
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.
Bae
Kwang-Hee
KH
ToolGen Inc., 461-6 Jeonmin-Dong, Yusung-Gu, Daejeon, 305-390, South Korea.
Kwon
Young Do
YD
Shin
Hyun-Chul
HC
Hwang
Moon-Sun
MS
Ryu
Eun-Hyun
EH
Park
Kyung-Soon
KS
Yang
Hyo-Young
HY
Lee
Dong-Ki
DK
Lee
Yangsoon
Y
Park
Jinwoo
J
Kwon
Heung Sun
HS
Kim
Hyun-Won
HW
Yeh
Byung-Il
BI
Lee
Hyean-Woo
HW
Sohn
Soon Hyung
SH
Yoon
Joonho
J
Seol
Wongi
W
Kim
Jin-Soo
JS
D004247:DNA / Q000737:chemistry / Q000235:genetics / Q000378:metabolism; D004268:DNA-Binding Proteins / Q000096:biosynthesis / Q000737:chemistry / Q000235:genetics; D005786:Gene Expression Regulation; D015894:Genome, Human; D006801:Humans; D019151:Peptide Library; D015202:Protein Engineering / Q000379:methods; D011994:Recombinant Proteins / Q000737:chemistry / Q000235:genetics / Q000378:metabolism; D014157:Transcription Factors / Q000096:biosynthesis* / Q000737:chemistry / Q000235:genetics*; D014158:Transcription, Genetic; D015003:Yeasts / Q000235:genetics / Q000378:metabolism; D016335:Zinc Fingers / Q000235:genetics*
D004268:DNA-Binding Proteins; D019151:Peptide Library; D011994:Recombinant Proteins; D014157:Transcription Factors; D004247:DNA