Tet-controlled binary transcription regulatory system is widely used in mammalian cells. Does anyone know if it is applicable to C elegans (in vivo and in cell lines)? If not, why?
Thanks
Tian
Dear Tian,
The answer is yes. The following publications cover the answer to your question.
1-Nat Methods. Author manuscript; available in PMC 2013 Dec 2.
Published in final edited form as:
Nat Methods. 2012 Mar 11; 9(4): 10.1038/nmeth.1929.
Published online 2012 Mar 11. doi: 10.1038/nmeth.1929
PMCID: PMC3846601
NIHMSID: NIHMS397900
Controlling gene expression with the Q repressible binary expression system inCaenorhabditis elegans
Xing Wei,1 Christopher J. Potter,1,2 Liqun Luo,1 and Kang Shen1
Author information ► Copyright and License information ►
The publisher's final edited version of this article is available at Nat Methods
See other articles in PMC that cite the published article.
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Abstract
We establish the first transcription-based binary gene expression system in C. elegans using the recently developed Q system. This system, derived from genes in Neurospora crassa, uses the transcriptional activator QF to induce the expression of target genes. Activation can be efficiently suppressed by the transcriptional repressor QS, and suppression in turn can be relieved by the non-toxic small molecule, quinic acid (QA). We used QF/QS and QA to achieve temporal and spatial control of transgene expression in various tissues in C. elegans. We further developed a Split Q system, in which we separated QF into two parts encoding its DNA-binding and transcription-activation domains. Each domain shows negligible transcriptional activity when expressed alone, but co-expression reconstitutes QF activity, providing additional combinatorial power to control gene expression.
2-Methods. Author manuscript; available in PMC 2015 Apr 1.
Published in final edited form as:
Methods. 2014 Apr 1; 66(3): 433–440.
Published online 2013 Jun 20. doi: 10.1016/j.ymeth.2013.06.012
PMCID: PMC3883888
NIHMSID: NIHMS527117
Adoption of the Q transcriptional regulatory system for zebrafish transgenesis
Abhignya Subedi,a,b Michelle Macurak,a Stephen T. Gee,c Estela Monge,a Mary G. Goll,a,1 Christopher J. Potter,dMichael J. Parsons,c and Marnie E. Halperna,*
Author information ► Copyright and License information ►
The publisher's final edited version of this article is available at Methods
See other articles in PMC that cite the published article.
Go to:
Abstract
The Gal4-UAS regulatory system of yeast is widely used to modulate gene expression in Drosophila; however, there are limitations to its usefulness in transgenic zebrafish, owing to progressive methylation and silencing of the CpG-rich multicopy upstream activation sequence. Although a modified, less repetitive UAS construct may overcome this problem, it is highly desirable to have additional transcriptional regulatory systems that can be applied independently or in combination with the Gal4/UAS system for intersectional gene expression. The Q transcriptional regulatory system of Neurospora crassa functions similarly to Gal4/UAS. QF is a transcriptional activator that binds to the QUAS upstream regulatory sequence to drive reporter gene expression. Unlike Gal4, the QF binding site does not contain essential CpG dinucleotide sequences that are subject to DNA methylation. The QS protein is a repressor of QF mediated transcriptional activation akin to Gal80. The functionality of the Q system has been demonstrated in Drosophila and C. elegans and we now report its successful application to a vertebrate model, the zebrafish, Danio rerio. Several tissue-specific promoters were used to drive QF expression in stable transgenic lines, as assessed by activation of a QUAS:GFP transgene. The QS repressor was found to dramatically reduce QF activity in injected zebrafish embryos; however, a similar repression has not yet been achieved in transgenic animals expressing QS under the control of ubiquitous promoters. A dual reporter construct containing both QUAS and UAS, each upstream of different fluorescent proteins was also generated and tested in transient assays, demonstrating that the two systems can work in parallel within the same cell. The adoption of the Q system should greatly increase the versatility and power of transgenic approaches for regulating gene expression in zebrafish.
Hoping this will be helpful,
Rafik
Dear Tian,
The answer is yes. The following publications cover the answer to your question.
1-Nat Methods. Author manuscript; available in PMC 2013 Dec 2.
Published in final edited form as:
Nat Methods. 2012 Mar 11; 9(4): 10.1038/nmeth.1929.
Published online 2012 Mar 11. doi: 10.1038/nmeth.1929
PMCID: PMC3846601
NIHMSID: NIHMS397900
Controlling gene expression with the Q repressible binary expression system inCaenorhabditis elegans
Xing Wei,1 Christopher J. Potter,1,2 Liqun Luo,1 and Kang Shen1
Author information ► Copyright and License information ►
The publisher's final edited version of this article is available at Nat Methods
See other articles in PMC that cite the published article.
Go to:
Abstract
We establish the first transcription-based binary gene expression system in C. elegans using the recently developed Q system. This system, derived from genes in Neurospora crassa, uses the transcriptional activator QF to induce the expression of target genes. Activation can be efficiently suppressed by the transcriptional repressor QS, and suppression in turn can be relieved by the non-toxic small molecule, quinic acid (QA). We used QF/QS and QA to achieve temporal and spatial control of transgene expression in various tissues in C. elegans. We further developed a Split Q system, in which we separated QF into two parts encoding its DNA-binding and transcription-activation domains. Each domain shows negligible transcriptional activity when expressed alone, but co-expression reconstitutes QF activity, providing additional combinatorial power to control gene expression.
2-Methods. Author manuscript; available in PMC 2015 Apr 1.
Published in final edited form as:
Methods. 2014 Apr 1; 66(3): 433–440.
Published online 2013 Jun 20. doi: 10.1016/j.ymeth.2013.06.012
PMCID: PMC3883888
NIHMSID: NIHMS527117
Adoption of the Q transcriptional regulatory system for zebrafish transgenesis
Abhignya Subedi,a,b Michelle Macurak,a Stephen T. Gee,c Estela Monge,a Mary G. Goll,a,1 Christopher J. Potter,dMichael J. Parsons,c and Marnie E. Halperna,*
Author information ► Copyright and License information ►
The publisher's final edited version of this article is available at Methods
See other articles in PMC that cite the published article.
Go to:
Abstract
The Gal4-UAS regulatory system of yeast is widely used to modulate gene expression in Drosophila; however, there are limitations to its usefulness in transgenic zebrafish, owing to progressive methylation and silencing of the CpG-rich multicopy upstream activation sequence. Although a modified, less repetitive UAS construct may overcome this problem, it is highly desirable to have additional transcriptional regulatory systems that can be applied independently or in combination with the Gal4/UAS system for intersectional gene expression. The Q transcriptional regulatory system of Neurospora crassa functions similarly to Gal4/UAS. QF is a transcriptional activator that binds to the QUAS upstream regulatory sequence to drive reporter gene expression. Unlike Gal4, the QF binding site does not contain essential CpG dinucleotide sequences that are subject to DNA methylation. The QS protein is a repressor of QF mediated transcriptional activation akin to Gal80. The functionality of the Q system has been demonstrated in Drosophila and C. elegans and we now report its successful application to a vertebrate model, the zebrafish, Danio rerio. Several tissue-specific promoters were used to drive QF expression in stable transgenic lines, as assessed by activation of a QUAS:GFP transgene. The QS repressor was found to dramatically reduce QF activity in injected zebrafish embryos; however, a similar repression has not yet been achieved in transgenic animals expressing QS under the control of ubiquitous promoters. A dual reporter construct containing both QUAS and UAS, each upstream of different fluorescent proteins was also generated and tested in transient assays, demonstrating that the two systems can work in parallel within the same cell. The adoption of the Q system should greatly increase the versatility and power of transgenic approaches for regulating gene expression in zebrafish.
Hoping this will be helpful,
Rafik
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