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Germline-competent stem cell in avian species and its application

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Asian Journal of Andrology 2015年第3期17卷 421-426页

作者： jae yong han Hyung Chul Lee Tae Sub Park Department of Agricultural Biotechnology College of Agriculture and Life Sciences and Research Institute of Agriculture and Life Sciences Seoul National University Seoul 151-921 Korea Graduate School of International Agricultural Technology and Institute of Green-Bio Science and Technology Seoul National University Pyeongchang-gun Gangwon-do 232-916 Korea

Germ cells are the only cell type in the body that can transfer genetic information to the next generation. Germline-competent stem cells can self-renew and contribute to the germ cell lineage giving rise to pluripotent stem cells under specific conditions. Hence far, studies on germline-competent stem cells have contributed to,the generation of avian model systems and the conservation of avian genetic resources. In this review, we focus on previous studies on germline-competent stem cells from avian species, mainly chicken germline-competent stem cells, which have been well established and characterized. We discuss different sources of germline-competent stem cells and recent advances for the future applications in birds.

关键词： birds germline-competent stem cells primordial germ cells spermatogonial stem cells

A modified,simple technique for internal limiting membrane free flap transplantation in macular hole surgery:the Peel-PFCL-Peel technique

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International Journal of Ophthalmology(English edition) 2021年第10期14卷 1636-1638页

作者： Hyun Goo Kang jae yong han jae Won Jun Min Kim Department of Ophthalmology Institute of Vision ResearchGangnam Severance HospitalYonsei University College of Medicine211 EonjuroGangnam-guSeoul 06273Republic of Korea

Dear Editor,We would like to introduce a simple but modified variant perfluorocarbon liquid(PFCL)-assisted technique,namely the"Peel-PFCL-Peel"technique,which overcomes these difficulties while improving manipulation ... 详细信息

关键词： modified limiting overcome

Differential transcriptional regulation of the NANOG gene in chicken primordial germ cells and embryonic stem cells

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Journal of Animal Science and Biotechnology 2021年第3期12卷 877-890页

作者： Hee Jung Choi So Dam Jin Deivendran Rengaraj Jin Hwa Kim Bertrand Pain jae yong han Department of Agricultural Biotechnology and Research Institute of Agriculture and Life Sciences Seoul National UniversitySeoul 08826South Korea Univ Lyon UniversitéLyon 1INSERMINRAEStem Cell and Brain Research InstituteU1208USC136169500 BronFrance Institute for Biomedical Sciences Shinshu UniversityMinamiminowaNagano 399-4598Japan

Background:NANOG is a core transcription factor(TF)in embryonic stem cells(ESCs)and primordial germ cells(PGCs).Regulation of the NANOG gene by TFs,epigenetic factors,and autoregulatory factors is well characterized in ESCs,and transcriptional regulation of NANOG is well established in these *** NANOG plays a key role in germ cells,the molecular mechanism underlying its transcriptional regulation in PGCs has not been ***,we investigated the mechanism that regulates transcription of the chicken NANOG(cNANOG)gene in PGCs and ***:We first identified the transcription start site of cNANOG by 5′-rapid amplification of cDNA ends PCR ***,we measured the promoter activity of various 5′flanking regions of cNANOG in chicken PGCs and ESCs using the luciferase reporter *** expression required transcriptional regulatory elements,which were positively regulated by POU5F3(OCT4)and SOX2 and negatively regulated by TP53 in *** proximal region of the cNANOG promoter contains a positive transcriptional regulatory element(CCAAT/enhancer-binding protein(CEBP)-binding site)in ***,small interfering RNA-mediated knockdown demonstrated that POU5F3,SOX2,and CEBP played a role in cell type-specific transcription of ***:We show for the first time that different trans-regulatory elements control transcription of cNANOG in a cell type-specific *** finding might help to elucidate the mechanism that regulates cNANOG expression in PGCs and ESCs.

关键词： Chicken Embryonic stem cells NANOG gene Primordial germ cells Regulatory elements

Sequential disruption of ALV host receptor genes reveals no sharing of receptors between ALV subgroups A, B, and J

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Journal of Animal Science and Biotechnology 2019年第2期10卷 410-423页

作者： Hong Jo Lee Kyung Je Park Kyung Youn Lee yongxiu Yao Venugopal Nair jae yong han Sciences and Research Institute of Agriculture and Life SciencesSeoul National UniversitySeoul 08826Korea The Pirbright Institute WokingSurrey GU24 0NFUK.

Background: Previously, we showed that targeted disruption of viral receptor genes in avian leukosis virus(ALV)subgroups using clustered regularly interspaced short palindromic repeats(CRISPR)/CRISPR-associated protein 9(Cas9))-based genome editing confers resistance to ALV subgroups B and J. Here, we used the same strategy to target the receptor expressed by ALV subgroup A(TVA) and generate chicken cells resistant to infection by this ***: CRISPR/Cas9-based disruption of exon 2 within the tva gene of DF-1 fibroblasts conferred resistance to infection by ALV subgroup A regardless of whether frameshift mutations were introduced during editing. Conversely,overexpression of the wild-type TVA receptor(wtTVA) by tva-modified DF-1 clones restored susceptibility to ALV subgroup A. The results confirm that exon 2, which contains the low-density lipoprotein receptor class A domain of TVA, is critical for virus entry. Furthermore, we sequentially modified DF-1 cells by editing the tva, tvb, and Na^+/H^+ exchange 1(chNHE1) genes, which are the specific receptors for ALV subgroups A, B, and J, ***: Simultaneous editing of multiple receptors to block infection by different subgroups of ALV confirmed that ALV subgroups A, B, and J do not share host receptors. This strategy could be used to generate cells resistant to multiple viral pathogens that use distinct receptors for cell entry.

关键词： Avian leukosis virus CRISPR/Cas9 Genome editing Host receptor TVA

Simple technique for the rescue and refixation of a partially disenclavated retropupillary iris claw intraocular lens

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International Journal of Ophthalmology(English edition) 2020年第11期13卷 1833-1835页

作者： Hyun Goo Kang jae yong han Sung Chul Lee Min Kim Department of Ophthalmology Institute of Vision ResearchGangnam Severance HospitalYonsei University College of Medicine211 EonjuroGangnam-guSeoul 06273Republic of Korea Department of Ophthalmology Institute of Vision ResearchSeverance Eye HospitalYonsei University College of Medicine50-1 Yonsei-roSeodaemun-guSeoul 03722Republic of Korea

Dear Editor,We would like to introduce a simple technique for the efficient rescue and refixation of a subluxated retropupillary iris claw intraocular lens(RPICIOL;Artisan■Aphakia model 205,Ophtec BV,Groningen,the Netherlands).The RPICIOL is a suture-less,posterior iris-fixated,polyinethyl methacrylate intraocular lens(IOL)that has gained widespread recognition and expanding use because of its efficiency in management of aphakia,combined with long-term intraocular safety and excellent sustained visual outcomes11 Postoperative dislocation/disenclavation of the RPICIOL may occur spontaneously or following trauma,with a rate reportedly between 0 to 10%[4-6].

关键词： law gained sustained

Single-cell RNA sequencing of mitoticarrested prospermatogonia with DAZL::GFP chickens and revealing unique epigenetic reprogramming of chickens

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Journal of Animal Science and Biotechnology 2022年第5期13卷 1305-1325页

作者： Hyeon Jeong Choi Kyung Min Jung Deivendran Rengaraj Kyung Youn Lee Eunhui Yoo Tae Hyun Kim jae yong han Department of Agricultural Biotechnology and Research Institute of Agriculture and Life Sciences Seoul National UniversitySeoul 08826South Korea Department of Animal Science Pennsylvania State UniversityState CollegePA 16801USA

Background:Germ cell mitotic arrest is conserved in many vertebrates,including birds,although the time of entry or exit into quiescence phase *** arrest is essential for the normal differentiation of male germ cells into spermatogonia and accompanies epigenetic reprogramming and meiosis inhibition from embryonic development to ***,mitotic arrest was not well studied in chickens because of the difficulty in obtaining pure germ cells from relevant developmental ***:We performed single-cell RNA sequencing to investigate transcriptional dynamics of male germ cells during mitotic arrest in DAZL::GFP *** differentially expressed gene analysis and K-means clustering to analyze cells at different developmental stages(E12,E16,and hatch),we found that metabolic and signaling pathways were regulated,and that the epigenome was reprogrammed during mitotic *** particular,we found that histone H3K9 and H3K14 acetylation(by HDAC2)and DNA demethylation(by DNMT3B and HELLS)led to a transcriptionally permissive chromatin ***,we found that global DNA demethylation occurred gradually after the onset of mitotic arrest,indicating that the epigenetic-reprogramming schedule of the chicken genome differs from that of the mammalian *** hypomethylation persisted after hatching,and methylation was slowly re-established 3 weeks ***:We found a unique epigenetic-reprogramming schedule of mitotic-arrested chicken prospermatogonia and prolonged hypomethylation after *** will provide a foundation for understanding the process of germ-cell epigenetic regulation in several species for which this process is not clearly *** findings on the biological processes related to sex-specific differentiation of prospermatogonia could help studying germline development in vitro more elaborately.

关键词： DAZL::GFP chickens DNA methylation Epigenetic reprogramming Histone acetylation Male germ cell Mitotic arrest Prospermatogonia Single-cell RNA sequencing

Regulatory elements and transcriptional control of chicken vasa homologue(CVH)promoter in chicken primordial germ cells

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Journal of Animal Science and Biotechnology 2017年第2期8卷 490-500页

作者： So Dam Jin Bo Ram Lee Young Sun Hwang Hong Jo Lee Jong Seop Rim jae yong han Department of Agricultural Biotechnology Research Institute of Agriculture and Life Sciences College of Agriculture and Life Sciences Seoul National University Institute for Biomedical Sciences Shinshu University

Background： Primordial germ cells（PGCs）, the precursors of functional gametes, have distinct characteristics and exhibit several unique molecular mechanisms to maintain pluripotency and germness in comparison to somatic cells. They express germ cel-specific RNA binding proteins（RBPs） by modulating tissue-specific cis-and trans-regulatory elements. Studies on gene structures of chicken vasa homologue（CVH）, a chicken RNA binding protein, involved in temporal and spatial regulation are thus important not only for understanding the molecular mechanisms that regulate germ cel fate, but also for practical applications of primordial germ cells. However, very limited studies are available on regulatory elements that control germ cel-specific expression in chicken. Therefore, we investigated the intricate regulatory mechanism（s） that governs transcriptional control of ***： We constructed green fluorescence protein（GFP） or luciferase reporter vectors containing the various 5′ flanking regions of CVH gene. From the 5′ deletion and fragmented assays in chicken PGCs, we have identified a CVH promoter that locates at-316 to ＋275 base pair fragment with the highest luciferase activity. Additional y, we confirmed for the first time that the 5′ untranslated region（UTR） containing intron 1 is required for promoter activity of the CVH gene in chicken PGCs. Furthermore, using a transcription factor binding prediction, transcriptome analysis and siR NA-mediated knockdown,we have identified that a set of transcription factors play a role in the PGC-specific CVH gene ***： These results demonstrate that cis-elements and transcription factors localizing in the 5′ flanking region including the 5′ UTR and an intron are important for transcriptional regulation of the CVH gene in chicken PGCs. Final y,this information wil contribute to research studies in areas of reproductive biology, constructing of germ cel-specific synthetic promoter for tracing primordial germ ce

关键词： Chicken Chicken vasa homologue Primordial germ cel Regulatory element siRNA-mediated knockdown

Acquisition of pluripotency in the chick embryo occurs during intrauterine embryonic development via a unique transcriptional network

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Journal of Animal Science and Biotechnology 2018年第3期9卷 574-584页

作者： jae yong han Hyo Gun Lee Young Hyun Park Young Sun Hwang Sang Kyung Kim Deivendran Rengaraj Byung Wook Cho Jeong Mook Lim Department of Agricultural Biotechnology and Research Institute of Agriculture and Life Sciences College of Agriculture and Life Sciences Seoul National University Institute for Biomedical Sciences Shinshu University Department of Animal Science and Technology Chung-Ang University Department of Animal Science College of Natural Resources and Life Sciences Pusan National University

Background: Acquisition of pluripotency by transcriptional regulatory factors is an initial developmental event that is required for regulation of cell fate and lineage specification during early embryonic development. The evolutionarily conserved core transcriptional factors regulating the pluripotency network in fishes, amphibians, and mammals have been elucidated. There are also species-specific maternally inherited transcriptional factors and their intricate transcriptional networks important in the acquisition of pluripotency. In avian species, however, the core transcriptional network that governs the acquisition of pluripotency during early embryonic development is not well ***: We found that chicken NANOG(c NANOG) was expressed in the stages between the pre-ovulatory follicle and oocyte and was continuously detected in Eyal-Giladi and Kochav stage I(EGK.I) to X. However, c POUV was not expressed during fol iculogenesis, but began to be detectable between EGK.V and VI. Unexpectedly, c SOX2 could not be detected during fol iculogenesis and intrauterine embryonic development. Instead of c SOX2, c SOX3 was maternally inherited and continuously expressed during chicken intrauterine development. In addition, we found that the pluripotency-related genes such as c ENS-1, c KIT, c LIN28 A, c MYC, c PRDM14, and c SALL4 began to be dramatical y upregulated between *** and ***: These results suggest that chickens have a unique pluripotent circuitry since maternally inherited c NANOG and c SOX3 may play an important role in the initial acquisition of pluripotency. Moreover, the acquisition of pluripotency in chicken embryos occurs at around *** to VI I.

关键词： Avian Embryonic development NANOG Pluripotency Transcriptional factor

Production of germline chimeric quails following spermatogonial cell transplantation in busulfan-treated testis

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Asian Journal of Andrology 2018年第4期20卷 414-416页

作者： Young Min Kim Jin Se Park Jong Won Yoon Hee Jung Choi Kyung Je Park Tamao Ono jae yong han Department of Agricultural Biotechnology Research Institute of Agriculture and Life Sciences College of Agriculture and Life Sciences Seoul National University Seou108826 Korea Division of Animal Science Faculty of Agriculture Shinshu University Minamiminowa Nagano 399-4598 Japan Institute for Biomedical Sciences Shinshu University Minamiminowa Nagano 399-4598 Japan

Dear Editor, The merits of using quaff as an avian experimental model include high egg production, low maintenance cost, small body size, and short generation period （approximately 6-8 weeks）. Indeed, these characteristics make quail an ideal species for many biological areas including transgenic research. Methods of primordial germ cell （PGC）-mediated germline chimera production are considered very reliable and have been used in avian transgenesis.

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