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Literature DB >> 9321676

A dicistronic construct allows easy detection of human CFTR expression from YAC DNA in human cells.

Abstract

We have made a dicistronic construct where the picornaviral internal ribosome-entry site (IRES) driving the expression of the beta-geo gene has been inserted into the 3'untranslated region of the human CFTR gene present in a YAC. When introduced into the human cell line Caco-2 expressing the CFTR gene, the expression of the dicistronic gene can be detected by lacZ staining and follows the accumulation of the endogenous CFTR mRNA upon differentiation of the cells. These data demonstrate that this IRES-based approach presents an alternative to mRNA in situ hybridisation and allows detection of expression in an autologous system.

Entities: Gene Species

Mesh：

Substances：

Year: 1997 PMID： 9321676 PMCID： PMC147010 DOI： 10.1093/nar/25.20.4167

Source DB: PubMed Journal: Nucleic Acids Res ISSN： 0305-1048 Impact factor: 16.971

14 in total

1. Targeting, disruption, replacement, and allele rescue: integrative DNA transformation in yeast.

Authors: R Rothstein
Journal: Methods Enzymol Date: 1991 Impact factor: 1.600

2. Complementation of null CF mice with a human CFTR YAC transgene.

Authors: A L Manson; A E Trezise; L J MacVinish; K D Kasschau; N Birchall; V Episkopou; G Vassaux; M J Evans; W H Colledge; A W Cuthbert; C Huxley
Journal: EMBO J Date: 1997-07-16 Impact factor: 11.598

3. Characterization of DNASE I hypersensitive sites in the 120kb 5' to the CFTR gene.

Authors: A N Smith; C J Wardle; A Harris
Journal: Biochem Biophys Res Commun Date: 1995-06-06 Impact factor: 3.575

4. A regulatory element in intron 1 of the cystic fibrosis transmembrane conductance regulator gene.

Authors: A N Smith; M L Barth; T L McDowell; D S Moulin; H N Nuthall; M A Hollingsworth; A Harris
Journal: J Biol Chem Date: 1996-04-26 Impact factor: 5.157

5. Use of yeast artificial chromosomes (YACs) for studying control of gene expression: correct regulation of the genes of a human beta-globin locus YAC following transfer to mouse erythroleukemia cell lines.

Authors: K R Peterson; G Zitnik; C Huxley; C H Lowrey; A Gnirke; K A Leppig; T Papayannopoulou; G Stamatoyannopoulos
Journal: Proc Natl Acad Sci U S A Date: 1993-12-01 Impact factor: 11.205

6. Dicistronic targeting constructs: reporters and modifiers of mammalian gene expression.

Authors: P Mountford; B Zevnik; A Düwel; J Nichols; M Li; C Dani; M Robertson; I Chambers; A Smith
Journal: Proc Natl Acad Sci U S A Date: 1994-05-10 Impact factor: 11.205

7. YAC transfer into mammalian cells by cell fusion.

Authors: N P Davies; C Huxley
Journal: Methods Mol Biol Date: 1996

8. Insertion of a pathogenic mutation into a yeast artificial chromosome containing the human amyloid precursor protein gene.

Authors: K Duff; A McGuigan; C Huxley; F Schulz; J Hardy
Journal: Gene Ther Date: 1994-01 Impact factor: 5.250

9. Use of yeast artificial chromosomes (YACs) in studies of mammalian development: production of beta-globin locus YAC mice carrying human globin developmental mutants.

Authors: K R Peterson; Q L Li; C H Clegg; T Furukawa; P A Navas; E J Norton; T G Kimbrough; G Stamatoyannopoulos
Journal: Proc Natl Acad Sci U S A Date: 1995-06-06 Impact factor: 11.205

A dicistronic construct allows easy detection of human CFTR expression from YAC DNA in human cells.

1. Targeting, disruption, replacement, and allele rescue: integrative DNA transformation in yeast.

2. Complementation of null CF mice with a human CFTR YAC transgene.

3. Characterization of DNASE I hypersensitive sites in the 120kb 5' to the CFTR gene.

4. A regulatory element in intron 1 of the cystic fibrosis transmembrane conductance regulator gene.

5. Use of yeast artificial chromosomes (YACs) for studying control of gene expression: correct regulation of the genes of a human beta-globin locus YAC following transfer to mouse erythroleukemia cell lines.

6. Dicistronic targeting constructs: reporters and modifiers of mammalian gene expression.

7. YAC transfer into mammalian cells by cell fusion.

8. Insertion of a pathogenic mutation into a yeast artificial chromosome containing the human amyloid precursor protein gene.

9. Use of yeast artificial chromosomes (YACs) in studies of mammalian development: production of beta-globin locus YAC mice carrying human globin developmental mutants.

10. Regulation of CFTR expression and function during differentiation of intestinal epithelial cells.

Review 1. Size matters: use of YACs, BACs and PACs in transgenic animals.

Review 2. Internal ribosome entry sites (IRESs): reality and use.

3. Regulated multicistronic expression technology for mammalian metabolic engineering.

4. Functional human CFTR produced by a stable minichromosome.