v

Chinese Livestock and Poultry Breeding ›› 2026, Vol. 22 ›› Issue (9): 95-102.doi: 10.19543/j.cnki.1673-4556.20260803.001cstr: 32418.14.j.cnki.1673-4556.20260803.001

• Regulation of Embryonic Development and Stem Cell Differentiation • Previous Articles     Next Articles

Research progress on the DLK1–DIO3 imprinted cluster in livestock embryonic development

Sihan Ju(), Liang Zhu, Wenqian Zhao, Zhonghua Liu, Xiaogang Weng()   

  1. College of Life Sciences, Northeast Agricultural University, Harbin, 150030, Heilongjiang
  • Received:2026-01-05 Online:2026-09-26 Published:2026-09-10
  • Contact: Xiaogang Weng

Abstract:

Imprinted genes are regulated through epigenetic mechanisms and are expressed in a parent-of-origin-specific manner. These genes are often clustered together and serve as crucial regulators in normal embryonic development and individual health. The DLK1-DIO3 imprinted cluster is a key regulatory region influencing livestock embryonic development and placental formation. The imprinting status of this cluster is primarily determined by the parent-of-origin-specific methylation of the core imprinting control region, IG-DMR(Imprinting Gene Differentially Methylated Region), which governs the overall parent-specific expression pattern of the cluster. GTL2-DMR(GTL2-Differentially Methylated Region), as a secondary DMR, maintains the stable expression of the maternal transcripts. This review summarizes the composition, imprinting regulation, and functional roles of this cluster, with a particular focus on the regulatory functions of the paternal genes DLK1, RTL1, DIO3, and the maternal gene GTL2 in placental angiogenesis, muscle growth, neural differentiation, and energy metabolism. The case of the Callipyge (CLPG) sheep, exhibiting a polar overdominant inheritance pattern, highlights the potential of manipulating this gene cluster to improve meat production traits and reproductive performance in livestock, so as to provide theoretical reference for improving the production performance and reproductive efficiency of livestock.

Key words: Genomic imprinting, DLK1-DIO3, Livestock, Embryonic development

CLC Number: 

  • S82

Fig. 1

Schematic diagram of the DLK1-DIO3 imprinted cluster Note: Blue and red represent the translation of paternally and maternally expressed genes, respectively, gray shapes represent imprinted genes with no expression."

Fig. 2

Regulatory model of the Callipyge phenotype Note: A→G:The CLPG mutation occurs in this region, where the original adenine (A) base is mutated to guanine (G). gray shapes represent imprinted genes with no expression.In the CLPG phenotype: CLPGPAT/+ animals exhibit overexpression of DLK1 leading to the CLPG phenotype; +/CLPGMAT animals display overexpression of maternal miRNAs and normal expression of paternal DLK1, which does not result in the CLPG phenotype; in CLPGPAT/CLPGMAT animals, paternal overexpression of DLK1 is post-transcriptionally suppressed by maternal overexpression of miRNAs, will not lead to CLPG phenotype."

[1]
WEINBERG-SHUKRON A, YOUNGSON N A, FERGUSON-SMITH A C, et al. Epigenetic control and genomic imprinting dynamics of the Dlk1-Dio3 domain[J]. Frontiers in Cell and Developmental Biology, 2023, 11: 1328806.
[2]
ALVES L F, SILVA I N DA, DE MELLO D C, et al. Epigenetic regulation of DLK1-DIO3 region in thyroid carcinoma[J]. Cells, 2024, 13(12): 1001.
[3]
VANNEVEL V, CLAES K, BAUD D, et al. Preeclampsia and long-term renal function in women who underwent kidney transplantation[J]. Obstetrics and Gynecology, 2018, 131(1): 57-62.
[4]
ZHANG X, HE H J, YU H R, et al. Maternal RNA transcription in Dlk1-Dio3 domain is critical for proper development of the mouse placental vasculature[J]. Communications Biology, 2024, 7: 363.
[5]
YU H R, ZHAO Y, CHENG R, et al. Silencing of maternally expressed RNAs in Dlk1-Dio3 domain causes fatal vascular injury in the fetal liver[J]. Cellular and Molecular Life Sciences, 2024, 81(1): 429.
[6]
HUBERT J N, PERRET M, RIQUET J, et al. Livestock species as emerging models for genomic imprinting[J]. Frontiers in Cell and Developmental Biology, 2024, 12: 1348036.
[7]
TAKADA S, TEVENDALE M, BAKER J, et al. Delta-like and Gtl2 are reciprocally expressed, differentially methylated linked imprinted genes on mouse chromosome 12[J]. Current Biology, 2000, 10(18): 1135-1138.
[8]
SATO S, YOSHIDA W, SOEJIMA H, et al. Methylation dynamics of IG-DMR and Gtl2-DMR during murine embryonic and placental development[J]. Genomics, 2011, 98(2): 120-127.
[9]
HELMAN L J, THIELE C J, LINEHAN W M, et al. Molecular markers of neuroendocrine development and evidence of environmental regulation[J]. Proceedings of the National Academy of Sciences of the United States of America, 1987, 84(8): 2336-2339.
[10]
LI M, PENG Y Q, SHI Y K, et al. Advancements in the study of DLK1 in the pathogenesis of diabetes[J]. Life Sciences, 2025, 369: 123535.
[11]
TASHIMA Y, TSUKAMOTO Y, TSUKAMOTO N, et al. Retention of DLK1 in the endoplasmic reticulum identifies roles for EGF-like-domain-specific O-glycans in the secretory pathway[J]. The FEBS Journal, 2025, 292(20): 5355-5381.
[12]
ESKICI N, GOMEZ-SANCHEZ C, MADHUSUDAN S, et al. Dual role of DLK1 in GnRH neuron ontogeny[J]. Stem Cell Reviews and Reports, 2025, 21(8): 2711-2726.
[13]
CHARLIER C, SEGERS K, WAGENAAR D, et al. Human-ovine comparative sequencing of a 250-kb imprinted domain encompassing the callipyge (clpg) locus and identification of six imprinted transcripts: DLK1, DAT, GTL2, PEG11, antiPEG11, and MEG8[J]. Genome Research, 2001, 11(5): 850-862.
[14]
ONO R, KOBAYASHI S, WAGATSUMA H, et al. A retrotransposon-derived gene, PEG10 is a novel imprinted gene located on human chromosome 7q21[J]. Genomics, 2001, 73(2): 232-237.
[15]
SHIURA H, KITAZAWA M, ISHINO F, et al. Roles of retrovirus-derived PEG10 and PEG11/RTL1 in mammalian development and evolution and their involvement in human disease[J]. Frontiers in Cell and Developmental Biology, 2023, 11: 1273638.
[16]
SHIURA H, KITAZAWA M, KANEKO-ISHINO T, et al. Placenta-driven evolution: viral gene acquisition and PEG10's essential roles in eutherian placenta[J]. Biomolecules, 2026, 16(1): 161.
[17]
SCHUSTER-GOSSLER K, SIMON-CHAZOTTES D, GUÉNET J L, et al. Gtl2lacZ, an insertional mutation on mouse Chromosome 12 with parental origin-dependent phenotype[J]. Mammalian Genome, 1996, 7(1): 20-24.
[18]
FARHADOVA S, GHOUSEIN A, CHARON F, et al. The long non-coding RNA Meg3 mediates imprinted gene expression during stem cell differentiation[J]. Nucleic Acids Research, 2024, 52(11): 6183-6200.
[19]
KAPLAN M M, YASKOSKI K A. Phenolic and tyrosyl ring deiodination of iodothyronines in rat brain homogenates[J]. The Journal of Clinical Investigation, 1980, 66(3): 551-562.
[20]
TSAI C E, LIN S P, ITO M, et al. Genomic imprinting contributes to thyroid hormone metabolism in the mouse embryo[J]. Current Biology, 2002, 12(14): 1221-1226.
[21]
TENG X Q, HE H J, YU H R, et al. LncRNAs in the Dlk1-Dio3 domain are essential for mid-embryonic heart development[J]. International Journal of Molecular Sciences, 2024, 25(15): 8184.
[22]
LI X J, YANG Y N, LI L, et al. Transcriptome profiling of different developmental stages on longissimus dorsi to identify genes underlying intramuscular fat content in wannanhua pigs[J]. Genes, 2023, 14(4): 903.
[23]
TRAUSTADÓTTIR G Á, JENSEN C H, THOMASSEN M, et al. Evidence of non-canonical NOTCH signaling: delta-like 1 homolog (DLK1) directly interacts with the NOTCH1 receptor in mammals[J]. Cellular Signalling, 2016, 28(4): 246-254.
[24]
HEJRATI N, LOU Z J, KOUHZAEI S, et al. DLK1-expressing neural progenitor cells promote tissue repair and functional recovery after cervical spinal cord injury[J]. Stem Cells Translational Medicine, 2025, 14(6): szaf014.
[25]
FERRÓN S R, CHARALAMBOUS M, RADFORD E, et al. Postnatal loss of Dlk1 imprinting in stem cells and niche astrocytes regulates neurogenesis[J]. Nature, 2011, 475(7356): 381-385.
[26]
SMITH C M, CATCHPOOLE D, HUTVAGNER G. MiRNAs from the Dlk1-Dio3 locus and miR-224/452 cluster contribute to glioblastoma tumor heterogeneity[J]. Scientific Reports, 2024, 14: 8570.
[27]
KITAZAWA M, HAYASHI S, IMAMURA M, et al. Deficiency and overexpression of Rtl1 in the mouse cause distinct muscle abnormalities related to Temple and Kagami-Ogata syndromes[J]. Development, 2020, 147(21): dev185918.
[28]
DAVIS E, CAIMENT F, TORDOIR X, et al. RNAi-mediated allelic trans-interaction at the imprinted Rtl1/Peg11 locus[J]. Current Biology, 2005, 15(8): 743-749.
[29]
ITO M, SFERRUZZI-PERRI A N, Edwards C A, et al. A trans-homologue interaction between reciprocally imprinted miR-127 and Rtl1 regulates placenta development[J]. Development, 2015: dev.121996.
[30]
田佺, 赵祎, 王卓, 等. 延边黄牛lncRNA MEG3基因多态性及其与生长性状的关联分析[J]. 中国畜牧兽医, 2021, 48(12): 4567-4574.
TIAN Q, ZHAO Y, WANG Z, et al. Polymorphisms of lncRNA MEG3 gene and its correlation with growth traits in Yanbian yellow cattle[J]. China Animal Husbandry Veterinary Medicine, 2021, 48(12): 4567-4574.
[31]
HERNANDEZ A, MARTINEZ M E, FIERING S, et al. Type 3 deiodinase is critical for the maturation and function of the thyroid axis[J]. The Journal of Clinical Investigation, 2006, 116(2): 476-484.
[32]
ISON E K, KENT-DENNIS C E, FAZIOLI J, et al. Compensatory mechanisms in response to induced hypothyroidism in the late gestation pig fetus[J]. Biology of Reproduction, 2023, 108(5): 731-743.
[33]
MARTINEZ M E, CHARALAMBOUS M, SAFERALI A, et al. Genomic imprinting variations in the mouse type 3 deiodinase gene between tissues and brain regions[J]. Molecular Endocrinology, 2014, 28(11): 1875-1886.
[34]
ARONSON B E, SCOURZIC L, SHAH V, et al. A bipartite element with allele-specific functions safeguards DNA methylation imprints at the Dlk1-Dio3 locus[J]. Developmental Cell, 2021, 56(22): 3052-3065.e5.
[35]
ZHANG L T, WANG J K, CAI G X, et al. Imprinted Dlk1-Gtl2 cluster miRNAs are potential epigenetic regulators of lamb fur quality[J]. BMC Genomics, 2023, 24(1): 632.
[36]
TAKEDA H, CAIMENT F, SMIT M, et al. The callipyge mutation enhances bidirectional long-range DLK1-GTL2 intergenic transcription in cis [J]. Proceedings of the National Academy of Sciences of the United States of America, 2006, 103(21): 8119-8124.
[37]
DAVIS E, JENSEN C H, SCHRODER H D, et al. Ectopic expression of DLK1 protein in skeletal muscle of padumnal heterozygotes causes the callipyge phenotype[J]. Current Biology, 2004, 14(20): 1858-1862.
[38]
TELLAM R L, COCKETT N E, VUOCOLO T, et al. Genes contributing to genetic variation of muscling in sheep[J]. Frontiers in Genetics, 2012, 3: 164.
[39]
FLEMING-WADDELL J N, OLBRICHT G R, TAXIS T M, et al. Effect of DLK1 and RTL1 but not MEG3 or MEG8 on muscle gene expression in Callipyge lambs[J]. PLoS One, 2009, 4(10): e7399.
[40]
GAO Y Q, CHEN X, WANG P, et al. Regulation of DLK1 by the maternally expressed miR-379/miR-544 cluster may underlie callipyge polar overdominance inheritance[J]. Proceedings of the National Academy of Sciences of the United States of America, 2015, 112(44): 13627-13632.
[41]
GUO J Z, ZHAO W, ZHAN S Y, et al. Identification and Expression Profiling of miRNAome in Goat longissimus dorsi Muscle from Prenatal Stages to a Neonatal Stage[J]. PLoS One, 2016, 11(10): e0165764.
[42]
王会, 柴志欣, 朱江江, 等. 牦牛Linc24063的克隆鉴定及其与miRNAs表达水平的相关性分析[J]. 中国农业科学, 2019, 52(14): 2538-2547.
WANG H, CHAI Z X, ZHU J J, et al. Cloning and identification of long-chain non-coding RNA Linc24063 and its correlation with the expression level of miRNAs in yak[J]. Scientia Agricultura Sinica, 2019, 52(14): 2538-2547.
[43]
RAZA S H A, KASTER N, KHAN R, et al. The role of microRNAs in muscle tissue development in beef cattle[J]. Genes, 2020, 11(3): 295.
[44]
LABIALLE S, MARTY V, BORTOLIN‐CAVAILLÉ M, et al. The miR‐379/miR‐410 cluster at the imprinted Dlk1‐Dio3 domain controls neonatal metabolic adaptation[J]. The EMBO Journal, 2014, 33(19): 2216-2230.
[45]
BOYSEN T J, TETENS J, THALLER G. Detection of a quantitative trait locus for ham weight with polar overdominance near the ortholog of the callipyge locus in an experimental pig F2 population[J]. Journal of Animal Science, 2010, 88(10): 3167-3172.
[46]
IMUMORIN I G, KIM E H, LEE Y M, et al. Genome scan for parent-of-origin QTL effects on bovine growth and carcass traits[J]. Frontiers in Genetics, 2011, 2: 44.
[47]
LI J L, YU D W, WANG J, et al. Identification of the porcine IG-DMR and abnormal imprinting of DLK1-DIO3 in cloned pigs[J]. Frontiers in Cell and Developmental Biology, 2022, 10: 964045.
[48]
FATHPOUR H, FOULADI M, JAFARPOUR F, et al. Crosstalk between myostatin and callipyge in CRISPR/Cas9-edited goat fibroblast cells[J]. Research in Veterinary Science, 2026, 198: 105992.
[49]
NI W, YOU S, CAO Y, et al. Aberrant expression of miR-127, miR-21 and miR-16 in placentas of deceased cloned sheep[J]. Research in Veterinary Science, 2016, 105: 200-204.
[50]
STEINHAUSER C B, ASKELSON K, HOBBS K C, et al. Maternal nutrient restriction alters thyroid hormone dynamics in placentae of sheep having small for gestational age fetuses[J]. Domestic Animal Endocrinology, 2021, 77: 106632.
[51]
LI Y H, XIAO P, BOADU F, et al. Beckwith-Wiedemann syndrome and large offspring syndrome involve alterations in methylome, transcriptome, and chromatin configuration[J]. medRxiv, 2025: 2023.12.14.23299981.
[52]
HARA S, MURAMATSU A, TERAO M, et al. Identification of maternal allele sequences of IG-DMR that are essential for neonatal viability[J]. PLoS One, 2025, 20(5): e0324882.
[53]
杨文志, 张明月, 王冠楠, 等. 2个印记的基因间lncRNAs位于牛Dlk1-Dio3印记区域[J]. 畜牧兽医学报, 2016, 47(9): 1848-1852.
YANG W Z, ZHANG M Y, WANG G N, et al. Two imprinted long non-coding RNAs located in cattle Dlk1-Dio3 domain[J]. Acta Veterinaria et Zootechnica Sinica, 2016, 47(9): 1848-1852.
[54]
杨莎, 郝海生, 杜卫华, 等. CRISPR/dCas9技术在基因表达调控中的研究进展[J]. 中国畜牧兽医, 2022, 49(1): 53-59.
YANG S, HAO H S, DU W H, et al. Research progress of CRISPR/dCas9 technology in gene expression regulation[J]. China Animal Husbandry Veterinary Medicine, 2022, 49(1): 53-59.
[55]
王丽娟. 正反交猪骨骼肌生长发育相关基因表达及表观遗传修饰的发育性变化研究[D]. 南京: 南京农业大学, 2011.
WANG, L J. Developmental changes of gene expression and epigenetic modifications related to skeletal muscle growth in reciprocal-cross pigs [D]. Nanjing: Nanjing Agricultural University, 2011.
[56]
张宸艺博, 余彤, 任斌斌, 等. 动物早期胚胎发育中表观重编程的机制[J]. 畜牧兽医学报, 2023, 54(12): 4898-4909.
ZHANG C, YU T, REN B B, et al. Mechanism of epigenetic reprogramming of early animal embryos[J]. Acta Veterinaria et Zootechnica Sinica, 2023, 54(12): 4898-4909.
[1] Bo Liu, Saizheng Han, Fangfang Sha, Wenwen Shen, Zihuai Tang, Weiwei Wu, Wenxin Zheng, Shiwei Zhou, Yinghui Wei, Xiaolong Wang. Challenges and prospects of generating cloned animals with livestock embryonic stem cells (ESCs) [J]. Chinese Livestock and Poultry Breeding, 2026, 22(9): 55-65.
[2] Jinying Zhang, Fan Zhao, xin Li, Suying Cao. The establishment and application prospects of epiblast stem cells in ruminant livestock [J]. Chinese Livestock and Poultry Breeding, 2026, 22(9): 47-54.
[3] Jinyun He, Zimo Zhao, Xiaowei Zhang, Zhiqiang Feng, Dengfeng Gao, Minglei Zhi. Advances in early embryonic development and embryo-derived stem cells in livestock [J]. Chinese Livestock and Poultry Breeding, 2026, 22(9): 12-25.
[4] Yu Shi, Liang Zhu, Sihan Ju, Zhonghua Liu, Xiaogang Weng. Advances in the regulation of lineage differentiation in early livestock embryos [J]. Chinese Livestock and Poultry Breeding, 2026, 22(9): 113-120.
[5] Qing Zhao, Zhiyan Zhao, Xueling Li. Research progress on the differentiation of domestic animal pluripotent stem cells into male germ cells [J]. Chinese Livestock and Poultry Breeding, 2026, 22(9): 103-112.
[6] Wanli Cheng, Mingyu Yang, Lei Xi, Zhuanjian Li, Yujie Guo. Research progress and prospects of intelligent measurement technologies for important phenotypes in livestock and poultry [J]. Chinese Livestock and Poultry Breeding, 2026, 22(7): 7-16.
[7] Ruiqi Cheng, Huaqian Zhou, Hua Yang, Yonglin Yang, Qian Yu, Wenzhe Zhang, Yan Chen, Zongsheng Zhao, lei Cui, Chunping Ma. Research progress on SNP chip development and its application in livestock and poultry genetic breeding [J]. Chinese Livestock and Poultry Breeding, 2026, 22(5): 61-69.
[8] Yuhao Cao, Yu Zou, Shijie Hu, Jiaqi Niu, Li Zhu, Ye Zhao, Mailin Gan. Relationship between intestinal dysbiosis and diarrhea in livestock and poultry, and related treatments [J]. Chinese Livestock and Poultry Breeding, 2026, 22(5): 21-31.
[9] Tiantian Wang, Jiahao Shao, Wenmiao Duan, Jianing Lu, Jianhua Zeng, Xiaohong Liu, Xiaolong Yuan. Research progress on genetic diversity of coat color in livestock genetic breeding [J]. Chinese Livestock and Poultry Breeding, 2026, 22(5): 108-115.
Viewed
Full text


Abstract

Cited

  Shared   
  Discussed   
No Suggested Reading articles found!