Improved applications of the tetracycline-regulated gene depletion system
- PMID: 20103842
Improved applications of the tetracycline-regulated gene depletion system
Abstract
Tightly controlled expression of transgenes in mammalian cells is an important tool for biological research, drug discovery, and future genetic therapies. The tetracycline-regulated gene depletion (Tet-Off) system has been widely used to control gene activities in mammalian cells, because it allows strict regulation of transgenes but no pleiotropic effects of prokaryotic regulatory proteins. However, the Tet-Off system is not compatible with every cell type and this is the main remaining obstacle left for this system. Recently, we overcame this problem by inserting an internal ribosome entry site (IRES) to drive a selectable marker from the same tetracycline-responsive promoter for the transgene. We also employed a CMV immediate early enhancer/beta-actin (CAG) promoter to express a Tet-controlled transactivator. Indeed, the Tet-Off system with these technical modifications was applied successfully to the human pre-B Nalm-6 cell line in which conventional Tet-Off systems had not worked efficiently. These methodological improvements should be applicable for many other mammalian proliferating cells. In this review we give an overview and introduce a new method for the improved application of the Tet-Off system.
Similar articles
-
Generation of tetracycline-inducible conditional gene knockout cells in a human Nalm-6 cell line.J Biotechnol. 2009 Apr 20;141(1-2):1-7. doi: 10.1016/j.jbiotec.2008.12.010. Epub 2008 Dec 24. J Biotechnol. 2009. PMID: 19135102
-
The tet-off system is more effective than the tet-on system for regulating transgene expression in a single adenovirus vector.J Gene Med. 2002 May-Jun;4(3):240-7. doi: 10.1002/jgm.261. J Gene Med. 2002. PMID: 12112641
-
Stringent and reproducible tetracycline-regulated transgene expression by site-specific insertion at chromosomal loci with pre-characterised induction characteristics.BMC Mol Biol. 2007 May 10;8:30. doi: 10.1186/1471-2199-8-30. BMC Mol Biol. 2007. PMID: 17493262 Free PMC article.
-
Tetracycline-regulated mouse models of cancer.Cold Spring Harb Protoc. 2014 Oct 1;2014(10):pdb.top069823. doi: 10.1101/pdb.top069823. Cold Spring Harb Protoc. 2014. PMID: 25275112 Review.
-
Tetracycline-controlled transcriptional regulation systems: advances and application in transgenic animal modeling.Semin Cell Dev Biol. 2002 Apr;13(2):121-8. doi: 10.1016/s1084-9521(02)00018-6. Semin Cell Dev Biol. 2002. PMID: 12127145 Review.
Cited by
-
DNA binding properties of the actin-related protein Arp8 and its role in DNA repair.PLoS One. 2014 Oct 9;9(10):e108354. doi: 10.1371/journal.pone.0108354. eCollection 2014. PLoS One. 2014. PMID: 25299602 Free PMC article.
-
Genetically engineered humanized mouse models for preclinical antibody studies.BioDrugs. 2014 Apr;28(2):171-80. doi: 10.1007/s40259-013-0071-0. BioDrugs. 2014. PMID: 24150980 Free PMC article. Review.
-
Neuronal transgene expression in dominant-negative SNARE mice.J Neurosci. 2014 Dec 10;34(50):16594-604. doi: 10.1523/JNEUROSCI.2585-14.2014. J Neurosci. 2014. PMID: 25505312 Free PMC article.
-
An improved Tet-on system in microRNA overexpression and CRISPR/Cas9-mediated gene editing.J Anim Sci Biotechnol. 2019 Jun 10;10:43. doi: 10.1186/s40104-019-0354-5. eCollection 2019. J Anim Sci Biotechnol. 2019. PMID: 31198556 Free PMC article.
-
Osterix-Cre transgene causes craniofacial bone development defect.Calcif Tissue Int. 2015 Feb;96(2):129-37. doi: 10.1007/s00223-014-9945-5. Epub 2014 Dec 31. Calcif Tissue Int. 2015. PMID: 25550101 Free PMC article.
Publication types
MeSH terms
Substances
LinkOut - more resources
Full Text Sources
Other Literature Sources
