2000
DOI: 10.1002/1526-968x(200009)28:1<31::aid-gene40>3.0.co;2-k
|View full text |Cite
|
Sign up to set email alerts
|

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

Abstract: A novel positive/negative selection cassette, puDeltatk, was generated. pu(Delta)tk is a bifunctional fusion protein between puromycin N-acetyltransferase (Puro) and a truncated version of herpes simplex virus type 1 thymidine kinase (DeltaTk). Murine embryonic stem (ES) cells transfected with pu(Delta)tk become resistant to puromycin and sensitive to 1-(-2-deoxy-2-fluoro-1-beta-D-arabino-furanosyl)-5-iodouracil (FIAU). Unlike other HSV1 tk transgenes, puDeltatk is readily transmitted through the male germ lin… Show more

Help me understand this report

Search citation statements

Order By: Relevance

Paper Sections

Select...
2
1
1
1

Citation Types

1
84
0

Year Published

2002
2002
2021
2021

Publication Types

Select...
7
1

Relationship

1
7

Authors

Journals

citations
Cited by 78 publications
(85 citation statements)
references
References 44 publications
1
84
0
Order By: Relevance
“…These enrichment strategies usually make use of negative selection markers such as the herpes simplex virus type 1 thymidine kinase gene or the diphtheria toxin ␣-chain coding region, which are placed outside the homology region of the targeting construct (10,20,161). Using a negative selection marker, ES cell clones with correct homologous recombination will not contain this marker, as removal of nonhomologous flanking sequences will take place prior to homologous integration.…”
Section: Conventional Germ Line Gene-targetingmentioning
confidence: 99%
See 1 more Smart Citation
“…These enrichment strategies usually make use of negative selection markers such as the herpes simplex virus type 1 thymidine kinase gene or the diphtheria toxin ␣-chain coding region, which are placed outside the homology region of the targeting construct (10,20,161). Using a negative selection marker, ES cell clones with correct homologous recombination will not contain this marker, as removal of nonhomologous flanking sequences will take place prior to homologous integration.…”
Section: Conventional Germ Line Gene-targetingmentioning
confidence: 99%
“…By contrast most randomly integrated targeting constructs still retain the negative selection marker, thus permitting counterselection against randomly integrated clones. In case of planning more complicated multistep genome engineering strategies, suitable selection markers allowing both positive and negative selection have also been developed (20).…”
Section: Conventional Germ Line Gene-targetingmentioning
confidence: 99%
“…To test whether the TW3R human feeder system can effectively support the selection and expansion of human iPSCs stably transfected with a piggyBac transposon, we used a piggyBac vector PB-CAG-GFP that co-expresses the GFP gene (under the CAG promoter) and Puro R -HSV-Dtk dual selection gene [33] (under the PGK promoter) to transfect BC1 human iPSCs (Fig. 5A).…”
Section: ‰ Engineered Human Feeders For Ipsc Derivation and Gene Targmentioning
confidence: 99%
“…5a). The marker gene consists of two parts linked head-to-tail: the first part encodes a fusion protein consisting of thymidine kinase and puromycin N-acetyltransferase 24 and the second encodes the target mRNA. Normally, ganciclovir kills cells carrying this marker gene owing to an intracellular accumulation of a toxic derivative of ganciclovir by thymidine kinase action 24 .…”
Section: Selection Of Efficient Shrna Constructs In Cellsmentioning
confidence: 99%
“…pMS240-PNS encodes the retrovirus vector expressing the thymidine kinase-puromycin-N-acetyltransferase fusion protein 24 under the control of the SV40 promoter. We constructed it from PCR-amplified fragments encoding thymidine kinase and puromycin acetyltransferase.…”
Section: Plasmid Constructionmentioning
confidence: 99%