Vrtn基因纯合敲除导致小鼠胚胎致死的多阶段转录组学分析

doi:10.16288/j.yczz.25-190

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Hereditas(Beijing) ›› 2026, Vol. 48 ›› Issue (3): 313-330.doi: 10.16288/j.yczz.25-190

• Research Article • Previous Articles Next Articles

Multi-stage transcriptomic analysis of mouse embryonic lethality induced by homozygous knockout of the Vrtn gene

Yaxin Deng¹(), Baojun Ding¹, Hongchun Li², Song Chen¹, Ying Zhang¹, Bo Zhou³(), Zhen Zhang¹^,⁴()

1. School of Basic Medical Sciences, Guangxi University Engineering Research Center for Rare Disease Prevention and Treatment, Guilin Medical University, Guilin 541199, China
2. School of Public Health, Guilin Medical University, Guilin 541199, China
3. Scientific Research Center, Guilin Medical University, Guilin 541199, China
4. Xiangya School of Basic Medical Sciences, Central South University, Changsha 410013, China

Received:2025-07-04 Revised:2025-10-27 Online:2026-03-20 Published:2025-11-21
Contact: Bo Zhou, Zhen Zhang E-mail:1528873272@qq.com;zb090900021@163.com;zhenzhang@glmu.edu.cn
Supported by:
National Natural Science Foundation of China(32060139);Natural Science Foundation of Guangxi Zhuang Autonomous Region(2025JJA130063);Natural Science Foundation of Guangxi Zhuang Autonomous Region(2020GXNSFBA297026);Graduate Research Program of Guilin Medical University(GYYK2025024);Guangxi Zhuang Autonomous Region Innovation and Entrepreneurship Training Program for College Students(S202510601101)

Abstract

Abstract:

The Vrtn gene, a recently identified regulator of embryonic development and stem cell pluripotency, is essential for embryonic survival, as its homozygous knockout (Vrtn^-/-) leads to lethality in mice at approximately embryonic day 12.5 (E12.5). To elucidate the underlying lethal mechanism, an integrated approach combining morphological observation, multi-stage transcriptomic analysis, and functional validation experiments was employed to systematically investigate the developmental disorders caused by Vrtn deficiency. Morphological observation showed that Vrtn^-/^- embryos exhibited significant abnormalities at developmental stages E9.0, E9.5, E10.0, E10.5, and E11.0, including shortened body axis, defective neural tube closure, aberrant somite differentiation, and cardiovascular malformations, accompanied by overall developmental delay. At the molecular level, through RNA sequencing and qRT-PCR validation revealed that Vrtn deficiency not only suppressed the expression of genes critical for somitogenesis (Hoxa2, Hes5), neurodevelopment (Nefm, Nefl), and the hematopoietic system (Hbb-bh1, Klf1), but also aberrantly activated genes associated with apoptosis (Crabp2, Fam162a) and lipid metabolism (Apom, Apoe). TUNEL staining showed that the level of apoptosis was significantly increased in Vrtn^-/- embryos. Meanwhile, immunofluorescence detection of Hif-1α indicated that the hypoxic stress response was aberrantly activated. Furthermore, the widespread dysregulation of genes involved in thyroid hormone transport (Ttr), DNA damage stress (Ddit4), and lipid metabolism (Apoa4, Apoa1) collectively exacerbated the developmental imbalance, ultimately leading to embryonic death. A cross-species analysis demonstrated that VRTN knockdown in human embryonic stem cells (hESCs) significantly suppressed the expression of core angiogenic genes (VEGFA, COL1A1 and HES1), a finding consistent with public database analyses indicating a strong association between VRTN and the hypoxic response. In conclusion, this study elucidates that Vrtn functions as a regulatory gene that maintains embryonic homeostasis by orchestrating multiple key developmental processes, including somitogenesis, neural differentiation, angiogenesis, and the hypoxic stress response. This discovery not only deepens the understanding of the role of Vrtn in embryonic development but also provides a new perspective for deciphering the pathogenesis of related hereditary diseases.

Key words: Vrtn gene, gene knockout, transcriptome, embryonic lethality

Yaxin Deng, Baojun Ding, Hongchun Li, Song Chen, Ying Zhang, Bo Zhou, Zhen Zhang. Multi-stage transcriptomic analysis of mouse embryonic lethality induced by homozygous knockout of the Vrtn gene[J]. Hereditas(Beijing), 2026, 48(3): 313-330.

Figures/Tables 9

Table 1

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Multi-stage transcriptomic analysis of mouse embryonic lethality induced by homozygous knockout of the Vrtn gene

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基因名称	引物序列(5′→3′)	扩增片段大小(bp)	退火温度(℃)
Hoxa2	F: TACGAATTTGAGCGAGAGATTGG	116	60
Hoxa2	R: GTCGAGGTCTTGATTGATGAACT
Hes5	F: AGTCCCAAGGAGAAAAACCGA	183	61
Hes5	R: GCTGTGTTTCAGGTAGCTGAC
Nefm	F: CTCGGCTATGCTCAGCTCG	96	59
Nefm	R: CGGGACAGTTTGTAGTCGCC
Nefl	F: CGCCGCTCTCAAGGACATC	129	60
Nefl	R: ACAGCGTCGGTGTTCTTGG
Hbb-bh1	F: GAAACCCCCGGATTAGAGCC	93	57
Hbb-bh1	R: GAGCAAAGGTCTCCTTGAGGT
Klf1	F: AGACTGTCTTACCCTCCATCAG	165	60
Klf1	R: GGTCCTCCGATTTCAGACTCAC
Vegfa	F: GCACATAGAGAGAATGAGCTTCC	105	61
Vegfa	R: CTCCGCTCTGAACAAGGCT
Bnip3	F: TCCTGGGTAGAACTGCACTTC	103	58
Bnip3	R: GCTGGGCATCCAACAGTATTT
Crabp2	F: ATGCCTAACTTTTCTGGCAACT	178	56
Crabp2	R: GCACAGTGGTGGAGGTTTTGA
Fam162a	F: AATGCTTCCTCGTCTCTAAGGT	117	61
Fam162a	R: CACCCCGTGTCTGTAAGACTG
Apom	F: TAACTCCATGAATCAGTGCCCT	105	60
Apom	R: CCCGCAATAAAGTACCACAGG
Apoe	F: CTGACAGGATGCCTAGCCG	107	57
Apoe	R: CGCAGGTAATCCCAGAAGC
Gapdh	F: AGGTCGGTGTGAACGGATTTG	123	60
Gapdh	R: TGTAGACCATGTAGTTGAGGTCA

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