Nuclear-envelope and nuclear-lamina binding chimeras for modulating gene expression

A technology for targeting genes and binding proteins, applied in the field of molecular switch systems for gene repression and derepression

Inactive Publication Date: 2005-05-18
先正达合作有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

[0006] However, known transcription factors have limited applications - such proteins are used to control genes related to their natural target sequences or to a limited set of closely related target sequences

Method used

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  • Nuclear-envelope and nuclear-lamina binding chimeras for modulating gene expression
  • Nuclear-envelope and nuclear-lamina binding chimeras for modulating gene expression
  • Nuclear-envelope and nuclear-lamina binding chimeras for modulating gene expression

Examples

Experimental program
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Embodiment 1

[0141] suppress human VEGF-A

[0142] To downregulate the expression of human vascular endothelial growth factor A (VEGF-A), a mouse GCL protein encoding 524 amino acids was prepared [Leatherman et al. (2000)] and the targeting sequence 5′-GTGTGG GTG AGT GAG TGT G-3 Recombinant construct of chimeric protein (CPI-vegf) of AZP' (SEQ ID NO: 7). A second construct encoding another chimeric protein (CP2-vegf) was made using the same mouse GCL protein and AZP targeting the sequence 5'-GGG GCT GGG GGCGGT GTC T-3' (SEQ ID NO: 8). The target nucleotide sequence is derived from the promoter of the human VEGF-A gene [Tischer et al. (1991) J. Biol. Chem. 266: 11947-11954]. The AZP has 6 zinc fingers, each with the framework sequence Pro-Tyr-Lys-Cys-Pro-Glu-Cys-Gly-Lys-Ser-Phe-Ser-Z -1 -Ser-Z 2 -Z 3 -Leu-Gln-Z 6 -His-Gln-Arg-Thr-His-Thr-Gly-Glu-Lys- (SEQ ID NO: 3); each frame is linked to the next with no redundant amino acid residues. Table 3 provides the binding specificity (Z -1 ...

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Abstract

The present invention relates to nucleic acid target-specific chimeric proteins comprising a nuclear envelope and / or lamina binding domain and a DNA binding domain. These proteins, as well as nucleic acids encoding these proteins, can be used in methods for suppressing or downregulating the expression of selected genes. The DNA binding domain is preferably derived from a naturally occurring zinc finger protein (ZFP) or an artificial zinc finger protein (AZP). The present invention also provides molecular switch systems for gene regulation.

Description

technical field [0001] The present invention relates to nucleic acid target-specific chimeric proteins comprising a nuclear-envelope and / or nuclear-lamina binding domain and a DNA binding domain. These proteins, as well as nucleic acids encoding these proteins, can be used in methods for modulating gene expression, in particular for repressing or downregulating the expression of selected target genes. The DNA binding domain is preferably derived from a naturally occurring zinc finger protein (ZFP) or an artificial zinc finger protein (AZP). The invention also relates to molecular switch systems for gene repression and derepression. Background technique [0002] Gene transcriptional repression can be accomplished through a variety of mechanisms. A classic example is the lac repressor, which, when bound to a target sequence on the lac operator, prevents RNA polymerase from binding and thus prevents initiation of transcription. In eukaryotes, other mechanisms exist to contro...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N15/09A61K35/76A61K38/00A61K48/00A61P9/00A61P25/30A61P31/20A61P35/00C07H21/04C07K14/00C07K14/47C07K19/00C12N1/15C12N1/19C12N1/21C12N5/02C12N5/10C12N9/22C12N15/62C12N15/63C12N15/82C12P21/02C12Q1/68
CPCC07K2319/09C07K2319/81C12N15/62C12N15/63C12N15/8217A61P25/30A61P31/20A61P35/00A61P9/00
Inventor 世良尚
Owner 先正达合作有限公司
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