scholarly journals Transient, Inducible, Placenta-Specific Gene Expression in Mice

Endocrinology ◽  
2012 ◽  
Vol 153 (11) ◽  
pp. 5637-5644 ◽  
Author(s):  
Xiujun Fan ◽  
Matthew Petitt ◽  
Matthew Gamboa ◽  
Mei Huang ◽  
Sabita Dhal ◽  
...  
Keyword(s):  
Gene Expression ◽  
Gene Delivery ◽  
Transgene Expression ◽  
Firefly Luciferase ◽  
Specific Gene ◽  
Dependent Manner ◽  
Control Of Gene Expression ◽  
Site Specific ◽  
Specific Gene Expression ◽  
High Level

Abstract Molecular understanding of placental functions and pregnancy disorders is limited by the absence of methods for placenta-specific gene manipulation. Although persistent placenta-specific gene expression has been achieved by lentivirus-based gene delivery methods, developmentally and physiologically important placental genes have highly stage-specific functions, requiring controllable, transient expression systems for functional analysis. Here, we describe an inducible, placenta-specific gene expression system that enables high-level, transient transgene expression and monitoring of gene expression by live bioluminescence imaging in mouse placenta at different stages of pregnancy. We used the third generation tetracycline-responsive tranactivator protein Tet-On 3G, with 10- to 100-fold increased sensitivity to doxycycline (Dox) compared with previous versions, enabling unusually sensitive on-off control of gene expression in vivo. Transgenic mice expressing Tet-On 3G were created using a new integrase-based, site-specific approach, yielding high-level transgene expression driven by a ubiquitous promoter. Blastocysts from these mice were transduced with the Tet-On 3G-response element promoter-driving firefly luciferase using lentivirus-mediated placenta-specific gene delivery and transferred into wild-type pseudopregnant recipients for placenta-specific, Dox-inducible gene expression. Systemic Dox administration at various time points during pregnancy led to transient, placenta-specific firefly luciferase expression as early as d 5 of pregnancy in a Dox dose-dependent manner. This system enables, for the first time, reliable pregnancy stage-specific induction of gene expression in the placenta and live monitoring of gene expression during pregnancy. It will be widely applicable to studies of both placental development and pregnancy, and the site-specific Tet-On G3 mouse will be valuable for studies in a broad range of tissues.

scholarly journals Site-specific gene expression analysis using an automated tissue micro-dissection punching system

2017 ◽  
Vol 7 (1) ◽  
Author(s):  
Takuya Yoda ◽  
Masahito Hosokawa ◽  
Kiyofumi Takahashi ◽  
Chikako Sakanashi ◽  
Haruko Takeyama ◽  
...  
Keyword(s):  
Gene Expression ◽  
Expression Analysis ◽  
Gene Expression Analysis ◽  
Specific Gene ◽  
Site Specific ◽  
Specific Gene Expression

Laser microdissection as a new tool to investigate site-specific gene expression in enteric ganglia of the human intestine

2010 ◽  
Vol 22 (2) ◽  
pp. 168-e52 ◽  
Author(s):  
m. böttner ◽  
f. bär ◽  
h. von koschitzky ◽  
k. tafazzoli ◽  
u. j. roblick ◽  
...  
Keyword(s):  
Gene Expression ◽  
Laser Microdissection ◽  
Specific Gene ◽  
Human Intestine ◽  
Site Specific ◽  
Specific Gene Expression ◽  
Enteric Ganglia

scholarly journals Dexamethasone-dependent inhibition of differentiation of C2 myoblasts bearing steroid-inducible N-ras oncogenes.

1988 ◽  
Vol 106 (6) ◽  
pp. 2127-2137 ◽  
Author(s):  
L A Gossett ◽  
W Zhang ◽  
E N Olson
Keyword(s):  
Gene Expression ◽  
Growth Factors ◽  
Specific Gene ◽  
Dependent Manner ◽  
Gene Products ◽  
Ras Proteins ◽  
Inhibitory Effects ◽  
Specific Gene Expression ◽  
Ras Oncogenes ◽  
Inhibition Of Differentiation

ras proteins are localized to the plasma membrane where they are postulated to interact with growth factor receptors and other proximal elements in intracellular cascades triggered by growth factors. The molecular events associated with terminal differentiation of certain skeletal myoblasts are inhibited by specific polypeptide growth factors and by constitutive expression of transforming ras oncogenes. To determine whether the inhibitory effects of ras on myogenic differentiation were reversible and to investigate whether muscle-specific genes remained susceptible to ras-dependent repression in terminally differentiated myotubes, the murine myoblast cell line, C2, was transfected with a plasmid containing a mutationally activated human N-ras oncogene under transcriptional control of the steroid-sensitive promoter of the mouse mammary tumor virus long terminal repeat. Addition of dexamethasone to myoblasts bearing steroid-inducible ras oncogenes prevented myotube formation and induction of muscle creatine kinase and acetylcholine receptors. Inhibition of differentiation by dexamethasone occurred in a dose-dependent manner and was a titratable function of ras expression. In the presence of dexamethasone, myoblasts bearing steroid-inducible ras genes retained their dependence on exogenous growth factors to divide and exhibited contact inhibition of growth at confluent densities, indicating that the inhibitory effects of ras on differentiation were independent of cell proliferation. Removal of dexamethasone from N-ras-transfected myoblasts led to fusion and induction of muscle-specific gene products in a manner indistinguishable from control C2 cells. Examination of the effects of culture media conditioned by ras-transfected myoblasts on differentiation of normal C2 cells yielded no evidence for inhibition of differentiation via an autocrine mechanism. In contrast to the ability of N-ras to prevent up-regulation of muscle-specific gene products in myoblasts, induction of N-ras in terminally differentiated myotubes failed to extinguish muscle-specific gene expression. Together, these results suggest that oncogenic ras proteins reversibly activate an intracellular cascade that prevents establishment of the differentiated phenotype. The inability of ras to extinguish muscle-specific gene expression in terminally differentiated myotubes also suggests that ras may interfere with an early step in the pathway of myoblasts toward the differentiated state.

Tu-P7:3 Sex- and site-specific gene expression in the adipose tissue of OB/OB mice

2006 ◽  
Vol 7 (3) ◽  
pp. 185
Author(s):  
S. Shiozaki ◽  
T. Chiba ◽  
K. Kokame ◽  
T. Miyata ◽  
M. Ai ◽  
...  
Keyword(s):  
Gene Expression ◽  
Adipose Tissue ◽  
Specific Gene ◽  
Site Specific ◽  
Specific Gene Expression

scholarly journals Intronic enhancers regulate the expression of genes involved in tissue-specific functions and homeostasis

2020 ◽  
Author(s):  
Beatrice Borsari ◽  
Pablo Villegas-Mirón ◽  
Hafid Laayouni ◽  
Alba Segarra-Casas ◽  
Jaume Bertranpetit ◽  
...  
Keyword(s):  
Gene Expression ◽  
Embryonic Stem ◽  
Gene Expression Pattern ◽  
Specific Gene ◽  
Control Of Gene Expression ◽  
Tissue Specific ◽  
Tissue Specific Gene ◽  
Specific Gene Expression ◽  
Expression Of Genes ◽  
Genomic Location

AbstractTissue function and homeostasis reflect the gene expression signature by which the combination of ubiquitous and tissue-specific genes contribute to the tissue maintenance and stimuli-responsive function. Enhancers are central to control this tissue-specific gene expression pattern. Here, we explore the correlation between the genomic location of enhancers and their role in tissue-specific gene expression. We found that enhancers showing tissue-specific activity are highly enriched in intronic regions and regulate the expression of genes involved in tissue-specific functions, while housekeeping genes are more often controlled by intergenic enhancers. Notably, an intergenic-to-intronic active enhancers continuum is observed in the transition from developmental to adult stages: the most differentiated tissues present higher rates of intronic enhancers, while the lowest rates are observed in embryonic stem cells. Altogether, our results suggest that the genomic location of active enhancers is key for the tissue-specific control of gene expression.

p130Cas-dependent actin remodelling regulates myogenic differentiation

2012 ◽  
Vol 445 (3) ◽  
pp. 323-332 ◽  
Author(s):  
Keiko Kawauchi ◽  
Wee Wee Tan ◽  
Keigo Araki ◽  
Farhana Binte Abu Bakar ◽  
Minsoo Kim ◽  
...  
Keyword(s):  
Gene Expression ◽  
Nuclear Localization ◽  
Myogenic Differentiation ◽  
Specific Gene ◽  
Control Of Gene Expression ◽  
Cytoskeletal Organization ◽  
Specific Gene Expression ◽  
Myotube Formation ◽  
Integrin Β3 ◽  
Actin Remodelling

Actin dynamics are implicated in various cellular processes, not only through the regulation of cytoskeletal organization, but also via the control of gene expression. In the present study we show that the Src family kinase substrate p130Cas (Cas is Crk-associated substrate) influences actin remodelling and concomitant muscle-specific gene expression, thereby regulating myogenic differentiation. In C2C12 myoblasts, silencing of p130Cas expression by RNA interference impaired F-actin (filamentous actin) formation and nuclear localization of the SRF (serum-response factor) co-activator MAL (megakaryocytic acute leukaemia) following the induction of myogenic differentiation. Consequently, formation of multinucleated myotubes was abolished. Re-introduction of wild-type p130Cas, but not its phosphorylation-defective mutant, into p130Cas-knockdown myoblasts restored F-actin assembly, MAL nuclear localization and myotube formation. Depletion of the adhesion molecule integrin β3, a key regulator of myogenic differentiation as well as actin cytoskeletal organization, attenuated p130Cas phosphorylation and MAL nuclear localization during C2C12 differentiation. Moreover, knockdown of p130Cas led to the activation of the F-actin-severing protein cofilin. The introduction of a dominant-negative mutant of cofilin into p130Cas-knockdown myoblasts restored muscle-specific gene expression and myotube formation. The results of the present study suggest that p130Cas phosphorylation, mediated by integrin β3, facilitates cofilin inactivation and promotes myogenic differentiation through modulating actin cytoskeleton remodelling.

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