Temporally and spatially controlled expression of transgenes in embryonic and adult tissues

Qian Zhang, Aleata A. Triplett, Don W. Harms, Wan Chi Lin, Bradley A. Creamer, A Angie Rizzino, Kay Uwe Wagner

Research output: Contribution to journalArticle

11 Citations (Scopus)

Abstract

Using ES cell-mediated transgenesis, we generated a novel mouse strain that permits a temporally and spatially controlled expression of responder genes in embryonic and multiple adult tissues. The transgene was constructed in a way that a CMV enhancer linked to the chicken β-actin promoter (CAG) drives the expression of the tetracycline-controlled transactivator (tTA) in particular tissues upon Cre-mediated excision of a floxed βgeo marker located between the promoter and the tTA. Based on the enzymatic activity of lacZ, the CAG-βgeo-tTA construct exhibits a widespread expression and appears to be very strong in the brain, heart, muscle, pancreas, and skin. Like the embryonic stem cell line that was used to generate this strain, the CAG-βgeo-tTA transgene is already highly active in preimplantation embryos. Using in vivo bioluminescence imaging on MMTV-Cre, CAG-βgeo-tTA, TetO-Luciferase triple transgenic mice and their controls, we demonstrated that the expression of the tTA, which is strictly dependent on the presence of Cre recombinase, induces the activation of the reporter transgene in the absence of any ligands. The tTA-mediated transactivation can be completely ablated through administration of doxycycline, and its subsequent withdrawal lifts the transcriptional block. Based on these characteristics, this novel strain may be useful in experiments that require a sustained expression of transgenes in particular cell types over a prolonged period followed by a rapid downregulation, for example in studies that examine the therapeutic value of cancer-initiating oncogenes during disease progression.

Original languageEnglish (US)
Pages (from-to)499-509
Number of pages11
JournalTransgenic Research
Volume19
Issue number3
DOIs
StatePublished - Jan 1 2010

Fingerprint

Trans-Activators
Transgenes
transgenes
promoter regions
Gene Transfer Techniques
doxycycline
bioluminescence
Doxycycline
oncogenes
embryonic stem cells
mice
Blastocyst
transcriptional activation
luciferase
Embryonic Stem Cells
tissues
transactivators
Tetracycline
Luciferases
tetracycline

Keywords

  • Cre recombinase
  • Gene expression regulation
  • Stem cells
  • Tetracycline
  • Trans-activators
  • Transgenic

ASJC Scopus subject areas

  • Biotechnology
  • Animal Science and Zoology
  • Agronomy and Crop Science
  • Genetics

Cite this

Zhang, Q., Triplett, A. A., Harms, D. W., Lin, W. C., Creamer, B. A., Rizzino, A. A., & Wagner, K. U. (2010). Temporally and spatially controlled expression of transgenes in embryonic and adult tissues. Transgenic Research, 19(3), 499-509. https://doi.org/10.1007/s11248-009-9329-1

Temporally and spatially controlled expression of transgenes in embryonic and adult tissues. / Zhang, Qian; Triplett, Aleata A.; Harms, Don W.; Lin, Wan Chi; Creamer, Bradley A.; Rizzino, A Angie; Wagner, Kay Uwe.

In: Transgenic Research, Vol. 19, No. 3, 01.01.2010, p. 499-509.

Research output: Contribution to journalArticle

Zhang, Q, Triplett, AA, Harms, DW, Lin, WC, Creamer, BA, Rizzino, AA & Wagner, KU 2010, 'Temporally and spatially controlled expression of transgenes in embryonic and adult tissues', Transgenic Research, vol. 19, no. 3, pp. 499-509. https://doi.org/10.1007/s11248-009-9329-1
Zhang, Qian ; Triplett, Aleata A. ; Harms, Don W. ; Lin, Wan Chi ; Creamer, Bradley A. ; Rizzino, A Angie ; Wagner, Kay Uwe. / Temporally and spatially controlled expression of transgenes in embryonic and adult tissues. In: Transgenic Research. 2010 ; Vol. 19, No. 3. pp. 499-509.
@article{3b3862d7ee85415593500c5f4171027d,
title = "Temporally and spatially controlled expression of transgenes in embryonic and adult tissues",
abstract = "Using ES cell-mediated transgenesis, we generated a novel mouse strain that permits a temporally and spatially controlled expression of responder genes in embryonic and multiple adult tissues. The transgene was constructed in a way that a CMV enhancer linked to the chicken β-actin promoter (CAG) drives the expression of the tetracycline-controlled transactivator (tTA) in particular tissues upon Cre-mediated excision of a floxed βgeo marker located between the promoter and the tTA. Based on the enzymatic activity of lacZ, the CAG-βgeo-tTA construct exhibits a widespread expression and appears to be very strong in the brain, heart, muscle, pancreas, and skin. Like the embryonic stem cell line that was used to generate this strain, the CAG-βgeo-tTA transgene is already highly active in preimplantation embryos. Using in vivo bioluminescence imaging on MMTV-Cre, CAG-βgeo-tTA, TetO-Luciferase triple transgenic mice and their controls, we demonstrated that the expression of the tTA, which is strictly dependent on the presence of Cre recombinase, induces the activation of the reporter transgene in the absence of any ligands. The tTA-mediated transactivation can be completely ablated through administration of doxycycline, and its subsequent withdrawal lifts the transcriptional block. Based on these characteristics, this novel strain may be useful in experiments that require a sustained expression of transgenes in particular cell types over a prolonged period followed by a rapid downregulation, for example in studies that examine the therapeutic value of cancer-initiating oncogenes during disease progression.",
keywords = "Cre recombinase, Gene expression regulation, Stem cells, Tetracycline, Trans-activators, Transgenic",
author = "Qian Zhang and Triplett, {Aleata A.} and Harms, {Don W.} and Lin, {Wan Chi} and Creamer, {Bradley A.} and Rizzino, {A Angie} and Wagner, {Kay Uwe}",
year = "2010",
month = "1",
day = "1",
doi = "10.1007/s11248-009-9329-1",
language = "English (US)",
volume = "19",
pages = "499--509",
journal = "Transgenic Research",
issn = "0962-8819",
publisher = "Springer Netherlands",
number = "3",

}

TY - JOUR

T1 - Temporally and spatially controlled expression of transgenes in embryonic and adult tissues

AU - Zhang, Qian

AU - Triplett, Aleata A.

AU - Harms, Don W.

AU - Lin, Wan Chi

AU - Creamer, Bradley A.

AU - Rizzino, A Angie

AU - Wagner, Kay Uwe

PY - 2010/1/1

Y1 - 2010/1/1

N2 - Using ES cell-mediated transgenesis, we generated a novel mouse strain that permits a temporally and spatially controlled expression of responder genes in embryonic and multiple adult tissues. The transgene was constructed in a way that a CMV enhancer linked to the chicken β-actin promoter (CAG) drives the expression of the tetracycline-controlled transactivator (tTA) in particular tissues upon Cre-mediated excision of a floxed βgeo marker located between the promoter and the tTA. Based on the enzymatic activity of lacZ, the CAG-βgeo-tTA construct exhibits a widespread expression and appears to be very strong in the brain, heart, muscle, pancreas, and skin. Like the embryonic stem cell line that was used to generate this strain, the CAG-βgeo-tTA transgene is already highly active in preimplantation embryos. Using in vivo bioluminescence imaging on MMTV-Cre, CAG-βgeo-tTA, TetO-Luciferase triple transgenic mice and their controls, we demonstrated that the expression of the tTA, which is strictly dependent on the presence of Cre recombinase, induces the activation of the reporter transgene in the absence of any ligands. The tTA-mediated transactivation can be completely ablated through administration of doxycycline, and its subsequent withdrawal lifts the transcriptional block. Based on these characteristics, this novel strain may be useful in experiments that require a sustained expression of transgenes in particular cell types over a prolonged period followed by a rapid downregulation, for example in studies that examine the therapeutic value of cancer-initiating oncogenes during disease progression.

AB - Using ES cell-mediated transgenesis, we generated a novel mouse strain that permits a temporally and spatially controlled expression of responder genes in embryonic and multiple adult tissues. The transgene was constructed in a way that a CMV enhancer linked to the chicken β-actin promoter (CAG) drives the expression of the tetracycline-controlled transactivator (tTA) in particular tissues upon Cre-mediated excision of a floxed βgeo marker located between the promoter and the tTA. Based on the enzymatic activity of lacZ, the CAG-βgeo-tTA construct exhibits a widespread expression and appears to be very strong in the brain, heart, muscle, pancreas, and skin. Like the embryonic stem cell line that was used to generate this strain, the CAG-βgeo-tTA transgene is already highly active in preimplantation embryos. Using in vivo bioluminescence imaging on MMTV-Cre, CAG-βgeo-tTA, TetO-Luciferase triple transgenic mice and their controls, we demonstrated that the expression of the tTA, which is strictly dependent on the presence of Cre recombinase, induces the activation of the reporter transgene in the absence of any ligands. The tTA-mediated transactivation can be completely ablated through administration of doxycycline, and its subsequent withdrawal lifts the transcriptional block. Based on these characteristics, this novel strain may be useful in experiments that require a sustained expression of transgenes in particular cell types over a prolonged period followed by a rapid downregulation, for example in studies that examine the therapeutic value of cancer-initiating oncogenes during disease progression.

KW - Cre recombinase

KW - Gene expression regulation

KW - Stem cells

KW - Tetracycline

KW - Trans-activators

KW - Transgenic

UR - http://www.scopus.com/inward/record.url?scp=77952236062&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=77952236062&partnerID=8YFLogxK

U2 - 10.1007/s11248-009-9329-1

DO - 10.1007/s11248-009-9329-1

M3 - Article

C2 - 19821046

AN - SCOPUS:77952236062

VL - 19

SP - 499

EP - 509

JO - Transgenic Research

JF - Transgenic Research

SN - 0962-8819

IS - 3

ER -