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Chromatin Immunoprecipitation Assay for Tissue-specific Genes using Early-stage Mouse Embryos

Chromatin Immunoprecipitation Assay for Tissue-specific Genes using Early-stage Mouse Embryos

Published on: April 29, 2011

The basic helix-loop-helix transcription factor Hand1 regulates mouse development as a homodimer.

Dong Hu¹, Ian C Scott, Fran Snider

¹Department of Biochemistry and Molecular Biology, Faculty of Medicine, University of Calgary, Calgary, AB, Canada; Department of Comparative Biology and Experimental Medicine, Faculty of Veterinary Medicine, University of Calgary, Calgary, AB, Canada; Center for Stem Cell Application and Research, Union Hospital, Tongji Medical College, Huazhong University of Science and Technology, Wuhan, China.

Developmental Biology

|August 6, 2013

Summary

Hand1 homodimer sufficiency is shown in mice, revealing its critical role in development. Dosage of Hand1 homodimer impacts survival, with implications for placental and cardiac development.

Keywords:

Basic helix-loop-helix Hand1 Homodimer Mouse development Transcription factor Trophoblast

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Chromatin Immunoprecipitation Assay for Tissue-specific Genes using Early-stage Mouse Embryos

Chromatin Immunoprecipitation Assay for Tissue-specific Genes using Early-stage Mouse Embryos

Published on: April 29, 2011

Expression Analysis of Mammalian Linker-histone Subtypes

Expression Analysis of Mammalian Linker-histone Subtypes

Published on: March 19, 2012

Area of Science:

Developmental biology
Molecular genetics
Genetics

Background:

Hand1, a basic helix-loop-helix transcription factor, is vital for embryonic development, including the placenta, yolk sac, and heart.
Hand1's role in trophoblast giant cell (TGC) differentiation is established, yet potential heterodimer partners are absent in TGCs.

Purpose of the Study:

To investigate the hypothesis that Hand1 functions as a homodimer during mouse development.
To assess the functional sufficiency and dosage effects of Hand1 homodimers in vivo.

Main Methods:

Generation of knock-in mice engineered to express a tethered Hand1 homodimer (Hand1(TH)).
Analysis of Hand1(TH) expression in homozygous (Hand1(TH/TH)) and heterozygous (Hand1(TH/-)) mice.
Evaluation of embryonic viability, placental development, and cardiac and craniofacial morphology.

Main Results:

Hand1(TH/-) conceptuses with only tethered Hand1 were viable and fertile, indicating homodimer sufficiency for survival.
A significant proportion of Hand1(TH/-) and all Hand1(TH/TH) embryos exhibited in utero lethality with severe placental defects and developmental abnormalities.
Dosage-dependent effects of Hand1(TH) were observed, with Hand1(TH/+) mice showing no adverse effects on viability or fertility.

Conclusions:

Hand1 homodimerization plays a dominant and essential role in mouse development.
The expression dosage of Hand1 homodimer is critical for embryonic survival.
Hand1 heterodimers may be dispensable or play a modulatory role in regulating Hand1 homodimer activity during development.