Bind-Predict
A Comprehensive resource for zinc containing DNA binding proteins


APPLICATIONS OF ZINC FINGER PROTEINS

1. RECOMBINATION:

2. ARTIFICIAL 'ZFP' INHIBITS DNA REPLICATION IN HUMAN PAPILOMAVIRUS :

3. ACTIVATION OF ENDOGENOUS GENES :

4. POLYDACTYL ZINC-FINGER PROTEINS - UNIQUE ADDRESSING WITHIN COMPLEX GENOMES:

5. ZFP TARGETED GENE REGULATION:


1. RECOMBINATION:

Restriction endonuclease domains are popular tools to engineered zinc finger domains and provide a means of directing nuclease activity to particular regions of DNA. The efficient homologous recombination of plasmids in vivo has been demonstrated by a zinc finger - FokI fusion injected into Xenopus laevis oocyte nuclei. The zinc finger - FokI fusion cuts the plasmid at specific site and activated the DNA molecules for recombination, which was 100 percent efficient in optimal conditions. Due to its high efficiency, it is hoped that such a procedure will allow the targeted recombination of desired DNA molecules for many in vivo applications.

Reference :

Michael Moore and Christopher Ullman, RECENT DEVELOPMENTS IN THE ENGINEERING OF ZINC FINGER PROTEINS, BRIEFINGS IN FUNCTIONAL GENOMICS AND PROTEOMICS., Vol No 4., Pages 342-355., January 2003.

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2. ARTIFICIAL 'ZFP' INHIBITS DNA REPLICATION IN HUMAN PAPILOMAVIRUS :

The AZP technology has been applied in the inhibition of replication of a mammalian DNA virus , human Papilomavirus (HPV) type 18. Two AZPs, designated AZP HPV- 1 and AZP HPV- 2, were designed to block binding of the HPV - 18 E2 replication protein to the replication region. Both the designed AZPs had much higher affinities towards the replication origin than did the E2 protein, and efficiently blocked E2 binding in vitro. In transient replication assay, both the AZPs inhibited the viral DNA replication : the AZPs HPV- 2, especially reduced the replication to approximately 10%.

Reference:

Takashi Mino, Tomoaki Mori, Naoki Matsumoto, Yusuke Mineta, Tomoyuki Okamoto, Yasuhiro Aoyama and Takashi Sera, APPLICATION OF ARTIFICIAL ZINC-FINGER TO INHIBITION OF DNA REPLICATION OF HUMAN PAPILOMAVIRUS, Nucleic Acids Symposium Series No.50, page no: 313-314.

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3. ACTIVATION OF ENDOGENOUS GENES :

The VEGF-A gene shows the presence of DNase I hypersensitive regions which are known to be more accessable to DNA binding proteins than the surrounding sequences and so may provide favorable target sequences. Three ZFPs targeted to these DNase I hypersensitive regions and fused to the VP16 or p65 activation domains activated the expression of VEGF-A between 2- and 15- folds.

Reference :

Michael Moore and Christopher Ullman, RECENT DEVELOPMENTS IN THE ENGINEERING OF ZINC FINGER PROTEINs, BRIEFINGS IN FUNCTIONAL GENOMICS AND PROTEOMICS. Vol No 4., Pages 342-355., January 2003.

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4. POLYDACTYL ZINC-FINGER PROTEINS - UNIQUE ADDRESSING WITHIN COMPLEX GENOMES :

Zinc- finger proteins which contains three zinc finger domains, bind to nine contiguous base pairs in a DNA molecule. Using structure based modeling a polypeptide linker was designed that fuses two three finger proteins which bind to 18 contiguous base pairs of DNA in a sequence-specific fashion. The expression of these proteins as fusion to the activation or repression domains allows transcription to be specifically up- or down- modulated within the human cells. Thus the polydactyl zinc-finger proteins can be applied as genome - specific transcriptional switches in gene-therapy strategies and the development of novel transgenic plants and animals.

Reference :

Qiang Liu, David J. Segal, Jayant B. Ghiara and Carlos F. Barbas III, DESIGN OF POLYDACTYL ZINC - FINGER PROTEINS FOR UNIQUE ADDRESSING WITHIN COMPLEX GENOMES, Proc. Natl. Acad. Sci. USA., Biochemistry, Vol. 94, pp. 5530, May 1997.

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5. ZFP TARGETED GENE REGULATION : GENOMEWIDE SINGLE GENE SPECIFICITY :

ZFP TF can repress target gene expression with single gene specificity. A ZFP TF repressor binds to an 18 base pair recognition sequence within the promoter region of the endogenos CHK2 gene and gives a > 10 - fold reduction in CHK2 mRNA and protein. This emphasizes the fact that ZFP TFs are presise tools for target validation and also highlights their potential as clinical therapeutics.

Reference :

Siyuan Tan, Dmitry Guschin, Albert Davalos, Ya-Li Lee, Andrew W. Snowden, Yann Jouvenot, H. Steven Zhang, Katherine Howes, Andrew R. McNamara, Albert Lai, Chris Ullman, Lindsey Reynolds, Michael Moore, Mark Isalan, Lutz-Peter Berg, Bradley Campos, Hong Qi, S. Kaye Spratt, Casey C. Case, Carl O. Pabo, Judith Campisi, and Philip D. Gregory, ZINC- FINGER PROTEIN TARGETED GENE REGULATION : GENOMEWIDE SINGLE-GENE SPECIFICITY, PNAS, October 14,2003, Vol.100, No.21, 11997-12002.

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