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Updated: May 23, 2026
Functional Manipulation of Maternal Gene Products Using In Vitro Oocyte Maturation in Zebrafish
Published on: April 22, 2017
Disruption of murine mp29/Syf2/Ntc31 gene results in embryonic lethality with aberrant checkpoint response
Chia-Hsin Chen1, Po-Chen Chu, Liekyeow Lee
1Institute of Biochemical Sciences, College of Life Science, National Taiwan University, Taipei, Taiwan.
Abstract:
Human p29 is a putative component of spliceosomes, but its role in pre-mRNA is elusive. By siRNA knockdown and stable overexpression, we demonstrated that human p29 is involved in DNA damage response and Fanconi anemia pathway in cultured cells. In this study, we generated p29 knockout mice (mp29(GT/GT)) using the mp29 gene trap embryonic stem cells to study the role of mp29 in DNA damage response in vivo. Interruption of mp29 at both alleles resulted in embryonic lethality. Embryonic abnormality occurred as early as E6.5 in mp29(GT/GT) mice accompanied with decreased mRNA levels of α-tubulin and Chk1. The reduction of α-tubulin and Chk1 mRNAs is likely due to an impaired post-transcriptional event. An aberrant G2/M checkpoint was found in mp29 gene trap embryos when exposed to aphidicolin and UV light. This embryonic lethality was rescued by crossing with mp29 transgenic mice. Additionally, the knockdown of zfp29 in zebrafish resulted in embryonic death at 72 hours of development postfertilization (hpf). A lower level of acetylated α-tubulin was also observed in zfp29 morphants. Together, these results illustrate an indispensable role of mp29 in DNA checkpoint response during embryonic development.
Insights
The gene p29 is essential for embryonic development and DNA damage response. Its absence causes embryonic lethality and checkpoint defects, highlighting its critical role in cell survival.
Area of Science:
- Molecular Biology
- Developmental Biology
- Genetics
Background:
- The function of human p29 in pre-mRNA splicing and DNA damage response remains unclear.
- Previous studies suggested a role for human p29 in DNA damage response and the Fanconi anemia pathway in cultured cells.
Purpose of the Study:
- To investigate the in vivo role of mouse p29 (mp29) in DNA damage response during embryonic development.
- To elucidate the function of mp29 using knockout mouse models.
Main Methods:
- Generation of mp29 knockout mice (mp29(GT/GT)) via gene trapping.
- Analysis of embryonic lethality, developmental abnormalities, and gene expression in mp29(GT/GT) embryos.
- Assessment of the G2/M cell cycle checkpoint in response to DNA damaging agents (aphidicolin, UV light).
- Rescue experiments using mp29 transgenic mice.
- Knockdown studies in zebrafish (zfp29) to assess developmental effects.
Main Results:
- mp29(GT/GT) mice exhibited embryonic lethality by E6.5.
- mp29(GT/GT) embryos showed decreased mRNA levels of α-tubulin and Chk1, suggesting impaired post-transcriptional regulation.
- Aberrant G2/M checkpoint response was observed in mp29 gene trap embryos upon exposure to DNA damaging agents.
- Embryonic lethality was rescued by introducing a functional mp29 transgene.
- Zebrafish knockdown of zfp29 resulted in embryonic death and reduced acetylated α-tubulin levels.
Conclusions:
- mp29 plays an indispensable role in the DNA damage response and checkpoint control during embryonic development.
- mp29 is crucial for maintaining genomic integrity and proper development, likely through post-transcriptional mechanisms affecting key proteins like α-tubulin and Chk1.
