ArticleNucleic acids research2009
Studying human telomerase gene transcription by a chromatinized reporter generated by recombinase-mediated targeting of a bacterial artificial chromosome.
Article in Nucleic acids research, 2009. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Cited by 12 papers.
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Who cites it
12 citing papers in PubMed, 14 citations in OpenAlex.
- The regulatory function of tandem repeat VNTR2-1 in hTERT gene involves basic Helix-loop-helix family transcription factors.Environmental and molecular mutagenesis · 2021Article
- Polymorphic tandem DNA repeats activate the human telomerase reverse transcriptase gene.Proceedings of the National Academy of Sciences of the United States of America · 2021Article
- Crispr/Cas9-mediated cleavages facilitate homologous recombination during genetic engineering of a large chromosomal region.Biotechnology and bioengineering · 2020Article
- ETS variant transcription factor 5 and c-Myc cooperate in derepressing the human telomerase gene promoter via composite ETS/E-box motifs.The Journal of biological chemistry · 2020Article
- Engineering a humanized telomerase reverse transcriptase gene in mouse embryonic stem cells.Scientific reports · 2019Article
- Telomerase gene expression bioassays indicate metabolic activation of genotoxic lower chlorinated polychlorinated biphenyls.Scientific reports · 2018Article
- Regulation of human and mouse telomerase genes by genomic contexts and transcription factors during embryonic stem cell differentiation.Scientific reports · 2017Article
- Repression of telomerase gene promoter requires human-specific genomic context and is mediated by multiple HDAC1-containing corepressor complexes.FASEB journal : official publication of the Federation of American Societies for Experimental Biology · 2017Article
- Human Telomerase Reverse Transcriptase (hTERT) Transcription Requires Sp1/Sp3 Binding to the Promoter and a Permissive Chromatin Environment.The Journal of biological chemistry · 2015Article
- Dual roles of c-Myc in the regulation of hTERT gene.Nucleic acids research · 2014Article
- A multi-step strategy for BAC recombineering of large DNA fragments.International journal of biochemistry and molecular biology · 2011Article
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Authors and funding
4 authors at 1 institution in 1 country.
Funding
Abstract
The endogenous human telomerase reverse transcriptase (hTERT) gene is repressed in somatic cells. To study the mechanisms of its repression, we developed a strategy of retrovirus-directed Cre recombinase-mediated BAC targeting, or RMBT, to generate single-copy integrations of BAC at pre-engineered chromosomal sites. This technique involved retroviral transduction of acceptor loci, containing an HSV thymidine kinase marker, and subsequent integration of BAC constructs into the acceptor sites, utilizing the loxP and lox511 sites present in the vector backbones. The BAC reporter, with a Renilla luciferase cassette inserted downstream of the hTERT promoter, was retrofitted with a puromycin marker. Through puromycin selection and ganciclovir counter-selection, a targeting efficiency of over 50% was achieved. We demonstrated that the activity and chromatin structures of the hTERT promoter in chromosomally integrated BAC reporter recapitulated its endogenous counterpart of the host cells. Therefore, we have established a genetically amendable platform to study chromatin and epigenetic regulation of the hTERT gene. The highly efficient and versatile RMBT technique has general applicability for studying largely unexplored chromatin-dependent mechanisms of promoter regulation of various genes.
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Registered trials
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