Modulation of gene activity by consecutive gene targeting of one creatine kinase M allele in mouse embryonic stem cells

Jan Van Deursen, R. Lovell-Badge, F. Oerlemans, J. Schepens, B. Wieringa

Research output: Contribution to journalArticle

35 Citations (Scopus)

Abstract

The cytosolic creatine kinases (CK's; EC 2.7.3.2.) BB, BM and MM are dimeric isoenzymes which have an important role in energy metabolism and display characteristic tissue- and stage-specific patterns of expression in mammals. To study the functional role of the distribution of the CK isoenzymes we have focussed on the modulation of expression of the genes encoding the individual B and M subunits, starting at the muscle creatine kinase (CKM) gene which is transcriptionally inactive during early embryogenesis. Using repeated rounds of gene targeting in mouse embryonic stem (ES) cells, two types of mutant cell lines were obtained. First, we generated a cell line in which insertion of a neomycin resistance (neo(r)) gene had disrupted one of the CKM alleles. Subsequently, from this cell line, following introduction of an insertion type vector designed for replacement of the muscle specific CKM-enhancer by the constitutively acting polyoma virus enhancer PyF441, several independent doubly targeted clones were isolated which all had insertions in the previously neo-disrupted CKM allele. In some of these ES clones, the targeted enhancer replacement resulted in gene correction and functional activation of the silent CKM gene. Dimerisation between the ectopically expressed CKM subunits and CKB subunits which are normally present at high levels in ES cells, led to the formation of the BM isoform of CK in these clones.

Original languageEnglish (US)
Pages (from-to)2637-2643
Number of pages7
JournalNucleic Acids Research
Volume19
Issue number10
StatePublished - 1991
Externally publishedYes

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MM Form Creatine Kinase
Gene Targeting
Alleles
Clone Cells
Cell Line
Isoenzymes
Genes
Polyomavirus
Neomycin
Dimerization
Embryonic Stem Cells
Creatine Kinase
Energy Metabolism
Embryonic Development
Mammals
Protein Isoforms
Gene Expression
Muscles
Mouse Embryonic Stem Cells

ASJC Scopus subject areas

  • Genetics

Cite this

Modulation of gene activity by consecutive gene targeting of one creatine kinase M allele in mouse embryonic stem cells. / Van Deursen, Jan; Lovell-Badge, R.; Oerlemans, F.; Schepens, J.; Wieringa, B.

In: Nucleic Acids Research, Vol. 19, No. 10, 1991, p. 2637-2643.

Research output: Contribution to journalArticle

Van Deursen, Jan ; Lovell-Badge, R. ; Oerlemans, F. ; Schepens, J. ; Wieringa, B. / Modulation of gene activity by consecutive gene targeting of one creatine kinase M allele in mouse embryonic stem cells. In: Nucleic Acids Research. 1991 ; Vol. 19, No. 10. pp. 2637-2643.
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