A new positive/negative selectable marker, puΔtk, for use in embryonic stem cells

genesis ◽  
2000 ◽  
Vol 28 (1) ◽  
pp. 31-35 ◽  
Author(s):  
You-Tzung Chen ◽  
Allan Bradley
Keyword(s):  
Stem Cells ◽  
Embryonic Stem Cells ◽  
Selectable Marker ◽  
Embryonic Stem ◽  
Negative Selectable Marker

scholarly journals Investigation of coelectroporation as a method for introducing small mutations into embryonic stem cells.

1992 ◽  
Vol 12 (6) ◽  
pp. 2769-2776 ◽  
Author(s):  
A C Davis ◽  
M Wims ◽  
A Bradley
Keyword(s):  
Stem Cells ◽  
Embryonic Stem Cells ◽  
Selectable Marker ◽  
Embryonic Stem ◽  
Chain Reaction ◽  
Genetic Changes ◽  
Replacement Vector ◽  
Hprt Locus ◽  
Polymerase Chain ◽  
Hypoxanthine Guanine Phosphoribosyltransferase

We have investigated coelectroporation as a method for introducing minor genetic changes into specific genes in embryonic stem cells. A selectable marker (neo) and a targeting replacement vector designed to insert a 4-bp insertion into exon 3 of the mouse hypoxanthine-guanine phosphoribosyltransferase (HPRT) gene were coelectroporated into embryonic stem cells and selected in G418 and 6-thioguanine (6-TG). HPRT-negative clones were obtained at a frequency of approximately 1 per 520 G418r clones. Southern analysis and the polymerase chain reaction were used to demonstrate that 3 of 36 of the 6-TG-resistant clones had the desired 4-bp insertion without any other disruption of the HPRT locus. Initial studies indicated that the other 33 6-TG-resistant clones probably resulted from the targeted integration of a concatemer containing both the targeting construct and the selectable neo gene.

scholarly journals Investigation of coelectroporation as a method for introducing small mutations into embryonic stem cells

1992 ◽  
Vol 12 (6) ◽  
pp. 2769-2776
Author(s):  
A C Davis ◽  
M Wims ◽  
A Bradley
Keyword(s):  
Stem Cells ◽  
Embryonic Stem Cells ◽  
Selectable Marker ◽  
Embryonic Stem ◽  
Chain Reaction ◽  
Genetic Changes ◽  
Replacement Vector ◽  
Hprt Locus ◽  
Polymerase Chain ◽  
Hypoxanthine Guanine Phosphoribosyltransferase

We have investigated coelectroporation as a method for introducing minor genetic changes into specific genes in embryonic stem cells. A selectable marker (neo) and a targeting replacement vector designed to insert a 4-bp insertion into exon 3 of the mouse hypoxanthine-guanine phosphoribosyltransferase (HPRT) gene were coelectroporated into embryonic stem cells and selected in G418 and 6-thioguanine (6-TG). HPRT-negative clones were obtained at a frequency of approximately 1 per 520 G418r clones. Southern analysis and the polymerase chain reaction were used to demonstrate that 3 of 36 of the 6-TG-resistant clones had the desired 4-bp insertion without any other disruption of the HPRT locus. Initial studies indicated that the other 33 6-TG-resistant clones probably resulted from the targeted integration of a concatemer containing both the targeting construct and the selectable neo gene.

A pgk::hprt fusion as a selectable marker for targeting of genes in mouse embryonic stem cells: disruption of the T-cell receptor δ-chain-encoding gene

Gene ◽  
1991 ◽  
Vol 105 (2) ◽  
pp. 263-267 ◽  
Author(s):  
Nathalie van der Lugt ◽  
Ets Robanus Maandag ◽  
Hein te Riele ◽  
Peter W. Laird ◽  
Anton Berns
Keyword(s):  
Stem Cells ◽  
T Cell ◽  
Embryonic Stem Cells ◽  
T Cell Receptor ◽  
Selectable Marker ◽  
Embryonic Stem ◽  
Cell Receptor ◽  
Mouse Embryonic Stem Cells ◽  
Encoding Gene ◽  
Δ Chain
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