Control of gene expression

a gene expression and control technology, applied in the field of gene expression control, can solve the problems of limited means, less progress in actual manipulation of gene expression to produce novel traits, and intervention may lead to a modulation of the level of eukaryotic gene expression

Inactive Publication Date: 2003-04-17
BENITEC AUSTRALIA +1
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Whilst recombinant DNA technology has provided significant progress in an understanding of the mechanisms regulating eukaryotic gene expression, much less progress has been made in the actual manipulation of gene expression to produce novel traits.
Moreover, there are only limited means by which human intervention may lead to a modulation of the level of eukaryotic gene expression.
The efficiency of both of these approaches in targeting the expression of specific genes is very low and highly variable results are usually obtained.
Accordingly, there currently exists no consensus as to the nature of genetic sequences which provide the most efficient means for repressing, delaying or otherwise reducing gene expression using existing technologies.
Moreover, such a high degree of variation exists between generations that it is not possible to predict the level of repression of a specific gene in the progeny of an organism in which gene expression was markedly modified.
However, such silencing of tandemly repeated gene copies is of little utility in an attempt to manipulate gene expression in an animal cell by recombinant means, wherein the sequences capable of targeting the expression of a particular gene are introduced at dispersed locati

Method used

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Examples

Experimental program
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example 2

[0274] Genetic Constructs Comprising BEV Polymerase Gene Sequences Linked to the CMV Promoter Sequence and / or the SV40L Promoter Sequence

[0275] 1. Commercial Plasmids

[0276] Plasmid pBluescriptII (SK+)

[0277] Plasmid pBluescriptII (SK+) is commercially available from Stratagene and comprises the LacZ promoter sequence and lacZ-alpha transcription terminator, with a multiple cloning site for the insertion of structural gene sequences therein. The plasmid further comprises the ColE1 and f1 origins of replication and ampicillin-resistance gene.

[0278] Plasmid pSVL

[0279] Plasmid pSVL is commercially-obtainable from Pharmacia and serves as a source of the SV40 late promoter sequence. The nucleotide sequence of pSVL is also publicly available as GenBank Accession Number U13868.

[0280] Plasmid pCR2.1

[0281] Plasmid pCR2.1 is commercially available from Invitrogen and comprises the LacZ promoter sequence and lacZ-.alpha. transcription terminator, with a cloning site for the insertion of structur...

example 3

[0346] Genetic constructs comprising the porcine .alpha.-1 ,3-galactosyltransferase (Gait) structural gene sequence or sequences operably connected to the CMV promoter sequence and / or the SV40L promoter sequence

[0347] 1. Commercial Plasmids

[0348] Plasmid pcDNA3

[0349] Plasmid pcDNA3 is commercially available from Invitrogen and comprises the CMV-lE promoter and BGHpA transcription terminator, with multiple cloning sites for the insertion of structural gene sequences there between. The plasmid further comprises the ColE1 and fl origins of replication and neomycin-resistance and ampicillin-resistance genes.

[0350] 2. Intermediate Plasmids

[0351] Plasmid pcDNA3.Galt

[0352] Plasmid pcDNA3.Galt (BresaGen Limited, South Australia, Australia; FIG. 28) is plasmid pcDNA3 (Invitrogen) and comprises the cDNA sequence encoding porcine gene alpha-1,3-galactosyltransferase (Galt) operably under the control of the CMV-lE promoter sequence such that it is capable of being expressed therefrom. To produc...

example 4

[0380] Genetic constructs comprising PVY Nia sequences operably linked to the35S promoter sequence and / or the SCBV promoter sequence

[0381] 1. Binary Vector

[0382] Plasmid pART27

[0383] Plasmid pART27 is a binary vector, specifically designed to be compatible with the pART7 expression cassette. It contains bacterial origins of replication for both E. coil and Agrobacterium tumefaciens, a spectinomycin resistance gene for bacterial selection, left and right T-DNA borders for transfer of DNA from Agrobacterium to plant cells and a kanamycin resistance cassette to permit selection of transformed plant cells. The kanamycin resistance cassette is located between the T-DNA borders, pART27 also contains a unique Notl restriction site which permits cloning of constructs prepared in vectors such as pART7 to be cloned between the T-DNA borders. Construction of pART27 is described in Gleave, AP (1992).

[0384] When cloning NotI inserts into this vector, two insert orientations can be obtained. In a...

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Abstract

The present invention relates generally to synthetic genes for modifying endogenous gene expression in a cell, tissue or organ of a transgenic organism, in particular a transgenic animal or plant. More particularly, the present invention provides novel synthetic genes and genetic constructs which are capable of repressing delaying or otherwise reducing the expression of an endogenous gene or a target gene in an organism when introduced thereto.

Description

[0001] This application is a continuation-in-part of U.S. Application Ser. No. 09 / 100,812.[0002] The present invention relates generally to a method of modifying gene expression and to synthetic genes for modifying endogenous gene expression in a call, tissue or organ of a transgenic organism, in particular a transgenic animal or plant. More particularly, the present invention utilises recombinant DNA technology to post-transcriptionally modify or modulate the expression of a target gene in a call, tissue, organ or whole organism, thereby producing novel phenotypes. Novel synthetic genes and genetic constructs which are capable of repressing delaying or otherwise reducing the expression of an endogenous gene or a target gene in an organism when introduced thereto are also provided.GENERAL[0003] Bibliographic details of the publications referred to in this specification are collected at the end of the description.[0004] As used herein the term "derived from" shall be taken to indicat...

Claims

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Application Information

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IPC IPC(8): C12N1/15A01K67/027C12N1/19C12N1/21C12N5/10C12N9/10C12N9/12C12N9/50C12N15/09C12N15/113C12N15/63C12N15/69C12N15/79C12N15/82C12N15/85C12R1/91
CPCA01K2217/05C12N2840/20C12N9/1051C12N9/127C12N9/503C12N15/111C12N15/113C12N15/63C12N15/69C12N15/8216C12N15/8218C12N15/8283C12N15/85C12N2310/111C12N2310/14C12N2310/531C12N2320/10C12N2320/30C12N2330/30C12N2330/50C12N2330/51C12N2800/108C12N2830/00C12N2830/002C12N2830/15C12N2830/38C12N2830/42C12N2830/55C12N2830/60A61K48/00C12N15/1131A61P31/04A61P31/12A61P43/00C12N15/10
Inventor GRAHAM, MICHAEL WAYNERICE, ROBERT NORMAN
Owner BENITEC AUSTRALIA
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