scholarly journals Development of Zinc Finger Domains for Recognition of the 5′-ANN-3′ Family of DNA Sequences and Their Use in the Construction of Artificial Transcription Factors

2001 ◽  
Vol 276 (31) ◽  
pp. 29466-29478 ◽  
Author(s):  
Birgit Dreier ◽  
Roger R. Beerli ◽  
David J. Segal ◽  
Jessica D. Flippin ◽  
Carlos F. Barbas
Keyword(s):  
Transcription Factors ◽  
Zinc Finger ◽  
Dna Sequences ◽  
Artificial Transcription Factors ◽  
Zinc Finger Domains

scholarly journals Development of Zinc Finger Domains for Recognition of the 5′-CNN-3′ Family DNA Sequences and Their Use in the Construction of Artificial Transcription Factors

2005 ◽  
Vol 280 (42) ◽  
pp. 35588-35597 ◽  
Author(s):  
Birgit Dreier ◽  
Roberta P. Fuller ◽  
David J. Segal ◽  
Caren V. Lund ◽  
Pilar Blancafort ◽  
...  
Keyword(s):  
Transcription Factors ◽  
Zinc Finger ◽  
Dna Sequences ◽  
Artificial Transcription Factors ◽  
Zinc Finger Domains

The reaction mechanism of FokI excludes the possibility of targeting zinc finger nucleases to unique DNA sites

2011 ◽  
Vol 39 (2) ◽  
pp. 584-588 ◽  
Author(s):  
Stephen E. Halford ◽  
Lucy E. Catto ◽  
Christian Pernstich ◽  
David A. Rusling ◽  
Kelly L. Sanders
Keyword(s):  
Zinc Finger ◽  
Dna Sequences ◽  
Catalytic Domain ◽  
Dna Recognition ◽  
Zinc Finger Nucleases ◽  
Target Sequence ◽  
Base Pairs ◽  
Catalytic Domains ◽  
Zinc Finger Domains ◽  
Better Than

The FokI endonuclease is a monomeric protein with discrete DNA-recognition and catalytic domains. The latter has only one active site so, to cut both strands, the catalytic domains from two monomers associate to form a dimer. The dimer involving a monomer at the recognition site and another from free solution is less stable than that from two proteins tethered to the same DNA. FokI thus cleaves DNA with two sites better than one-site DNA. The two sites can be immediately adjacent, but they can alternatively be many hundreds of base pairs apart, in either inverted or repeated orientations. The catalytic domain of FokI is often a component of zinc finger nucleases. Typically, the zinc finger domains of two such nucleases are designed to recognize two neighbouring DNA sequences, with the objective of cutting the DNA exclusively between the target sequences. However, this strategy fails to take account of the fact that the catalytic domains of FokI can dimerize across distant sites or even at a solitary site. Additional copies of either target sequence elsewhere in the chromosome must elicit off-target cleavages.

scholarly journals Phenotypic Alteration and Target Gene Identification Using Combinatorial Libraries of Zinc Finger Proteins in Prokaryotic Cells

2005 ◽  
Vol 187 (15) ◽  
pp. 5496-5499 ◽  
Author(s):  
Kyung-Soon Park ◽  
Young-Soon Jang ◽  
Horim Lee ◽  
Jin-Soo Kim
Keyword(s):  
Transcription Factors ◽  
Zinc Finger ◽  
Chromatin Immunoprecipitation ◽  
Target Gene ◽  
Combinatorial Libraries ◽  
Artificial Transcription Factors ◽  
Phenotypic Alteration ◽  
Target Dna ◽  
Genomic Studies ◽  
Phenotypic Variations

ABSTRACT We have developed a method with prokaryotic organisms that uses randomized libraries of zinc finger-containing artificial transcription factors to induce phenotypic variations and to identify genes involved in the generation of a specific phenotype of interest. Combining chromatin immunoprecipitation experiments and in silico prediction of target DNA binding sequences for the artificial transcription factors, we identified ubiX, whose down-regulation correlates with the thermotolerance phenotype in Escherichia coli. Our results show that randomized libraries of artificial transcription factors are powerful tools for functional genomic studies.

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