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Primordial germ cell-mediated transgenesis and genome editing in birds

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Journal of Animal Science and Biotechnology 2018年第2期9卷 257-267页

作者： Jae Yong Han Young Hyun Park 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

Transgenesis and genome editing in birds are based on a unique germline transmission system using primordial germ cells(PGCs), which is quite different from the mammalian transgenic and genome editing system. PGCs are progenitor cells of gametes that can deliver genetic information to the next generation. Since avian PGCs were first discovered in nineteenth century, there have been numerous efforts to reveal their origin, specification, and unique migration pattern, and to improve germline transmission efficiency. Recent advances in the isolation and in vitro culture of avian PGCs with genetic manipulation and genome editing tools enable the development of valuable avian models that were unavailable before. However, many challenges remain in the production of transgenic and genome-edited birds,including the precise control of germline transmission, introduction of exogenous genes, and genome editing in ***, establishing reliable germline-competent PGCs and applying precise genome editing systems are critical current issues in the production of avian models. Here, we introduce a historical overview of avian PGCs and their application, including improved techniques and methodologies in the production of transgenic and genome-edited birds, and we discuss the future potential applications of transgenic and genome-edited birds to provide opportunities and benefits for humans.

关键词： Avian Genome editing Primordial germ cell Transgenesis

Efficient generation of zebrafish maternal-zygotic mutants through transplantation of ectopically induced and Cas9/gRNA targeted primordial germ cells

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Journal of Genetics and Genomics 2020年第1期47卷 37-47页

作者： Fenghua Zhang Xianmei Li Mudan He Ding Ye Feng Xiong Golpour Amin Zuoyan Zhu Yonghua Sun State Key Laboratory of Freshwater Ecology and Biotechnology Innovation Academy for Seed DesignInstitute of HydrobiologyChinese Academy of SciencesWuhan430072China College o f Advanced Agricultural Sciences University of Chinese Academy of SciencesBeijing100049China

The clustered regularly interspaced short palindromic repeats(CRISPR)/Cas9 technology has been widely utilized for knocking out genes involved in various biological processes in zebrafish. Despite this technology is efficient for generating different mutations, one of the main drawbacks is low survival rate during embryogenesis when knocking out some embryonic lethal genes. To overcome this problem, we developed a novel strategy using a combination of CRISPR/Cas9 mediated gene knockout with primordial germ cell(PGC) transplantation(PGCT) to facilitate and speed up the process of zebrafish mutant generation, particularly for embryonic lethal genes. Firstly, we optimized the procedure for CRISPR/Cas9 targeted PGCT by increasing the efficiencies of genome mutation in PGCs and induction of PGC fates in donor embryos for PGCT. Secondly, the optimized CRISPR/Cas9 targeted PGCT was utilized for generation of maternal-zygotic(MZ) mutants of tcf7l1a(gene essential for head development), pou5f3(gene essential for zygotic genome activation) and chd(gene essential for dorsal development) at F1 generation with relatively high efficiency. Finally, we revealed some novel phenotypes in MZ mutants of tcf7l1 a and chd, as MZtcf7l1 a showed elevated neural crest development while MZchd had much severer ventralization than its zygotic counterparts. Therefore, this study presents an efficient and powerful method for generating MZ mutants of embryonic lethal genes in zebrafish. It is also feasible to speed up the genome editing in commercial fishes by utilizing a similar approach by surrogate production of CRISPR/Cas9 targeted germ cells.

关键词： Zebrafish CRISPR/Cas9 Primordial germ cells Transplantation Maternal zygotic mutant

In vitro reconstitution of germ cell development

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Protein & Cell 2011年第12期2卷 944-945页

作者： Yun Li 机构不详

Mammalian germ line cells undergo unique cellular and genetic changes under the regulation of specific regulators,in different stages as they develop and differentiate into functional *** is a fundamental challenge to reconstitute gametes development in vitro,because of the complexity of the regulation *** mice,embryonic stem cells(ESCs)develop into Epiblast stem cells at around embryonic day 6.0(E6.0)induced by the bone morphogenetic protein 4(Bmp4)(Lawson et al.,1999).At around E7.25,epiblast stem cells develop into primordial germ cells(PGCs)in the extraembryonic mesoderm regulated by the critical transcriptional regulator Blimp1(Prdm1)and Prdm14(Yamaji et al.,2008).PGCs are the origins for the oocytes and the spermatozoa(a motile sperm cell).

关键词： al. process. germ

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Fish germ cells

Science China(Life Sciences) 2010年第4期53卷 435-446页

作者： XU HongYan1, LI MingYou1, GUI JianFang2 & HONG YunHan1,2 1Department of Biological Sciences, National University of Singapore, Singapore 119260, Singapore 2 State Key Laboratory of Freshwater Ecology and Biotechnology, Institute of Hydrobiology, Chinese Academy of Sciences, Wuhan 430072, China 1. Department of Biological Sciences National University of Singapore Singapore 119260 Singapore2. State Key Laboratory of Freshwater Ecology and Biotechnology Institute of Hydrobiology Chinese Academy of Sciences Wuhan 430072 China

Fish, like many other animals, have two major cell lineages, namely the germline and soma. The germ-soma separation is one of the earliest events of embryonic development. germ cells can be specifically labeled and isolated for culture and transplan-tation, providing tools for reproduction of endangered species in close relatives, such as surrogate production of trout in salmon. Haploid cell cultures, such as medaka haploid embryonic stem cells have recently been obtained, which are capable of mimicking sperm to produce fertile offspring, upon nuclear being directly transferred into normal eggs. Such fish originated from a mosaic oocyte that had a haploid meiotic nucleus and a transplanted haploid mitotic cell culture nucleus. The first semi-cloned fish is Holly. Here we review the current status and future directions of understanding and manipulating fish germ cells in basic research and reproductive technology.

关键词： fish germ cell germ plasm reproduction reproductive technology semi-cloning

Association between KITLG Gene Polymorphisms and Testicular germ Cell Tumors:A Systematic Review and Meta-analysis

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Current Medical Science 2019年第5期39卷 702-706页

作者： Jia-xuan QIN Zong-long CAI Yu-feng YANG Jin-chun XING Xuan ZHUANG Department of Urology Surgery Center of Diagnosis and Treatment of Urinary System Diseasesthe Key Laboratory of Urinary Tract Tumors and Calculi of Xiamen Citythe First Affiliated Hospital of Xiamen UniversityXiamen 361003China The First Clinical Medical School of Fujian Medical University Xiamen 361003China

It has been reported that c-KIT ligand(KITLG)gene polymorphisms may be associated with testicular germ cell tumors(TGCT).Owing to mixed and inconclusive results,we con ducted a systematic review and meta-analysis to summarize and clarify this association.A systematic search of studies on the association between KITLG gene polymorphisms and TGCT susceptibility was conducted in *** ratios and 95%confidence intervals were used to pool the effect *** articles were included in our systematic review and *** with adenine(A),KITLG rs995030 guanine(G)might be associated with increased risk of *** are insufficient data to fully confirm the association between KITLG rs4474514 and TGCT ***-designed studies with larger sample size and more subgroups are required to validate the risk identified in the current meta-analysis.

关键词： c-KIT ligand single nucleotide polymorphisms testicular germ cell tumors metaanalysis

Reprogramming of germ cells into pluripotency

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World Journal of Stem Cells 2016年第8期8卷 251-259页

作者： Yoichi Sekita Toshinobu Nakamura Tohru Kimura Laboratory of Stem Cell Biology Department of Biosciences Kitasato University School of Science Laboratory of Epigenetic Regulation Department of Animal Bioscience Nagahama Institute of Bio-Science and Technology

Primordial germ cells(PGCs) are precursors of all gametes, and represent the founder cells of the germline. Although developmental potency is restricted to germ-lineage cells, PGCs can be reprogrammed into a pluripotent state. Specifically, PGCs give rise to germ cell tumors, such as testicular teratomas, in vivo, and to pluripotent stem cells known as embryonic germ cells in vitro. In this review, we highlight the current knowledge on signaling pathways, transcriptional controls, and post-transcriptional controls that govern germ cell differentiation and de-differentiation. These regulatory processes are common in the reprogramming of germ cells and somatic cells, and play a role in the pathogenesis of human germ cell tumors.

关键词： Primordial germ cell Embryonic germ cell germ cell tumor Reprogramming Induced pluripotent stem cell Small molecule compound Gene Signal Transcription factor

Retroperitoneal germ cell tumor diagnosed by endoscopic ultrasound-guided fine needle aspiration

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World Journal of Gastrointestinal Oncology 2010年第12期2卷 443-445页

作者： Craig M Womeldorph Mark M Zalupski Stewart M Knoepp Mohammad Soltani B Joseph Elmunzer Division of Gastroenterology University of Michigan Medical CenterAnn ArborMI 48109United States Division of Hematology and Oncology University of Michigan Medical CenterAnn ArborMI 48109United States Department of Pathology University of Michigan Medical CenteAnn ArborMI 48109United States

Isolated extragonadal germ cell tumors can be primary in nature or metastatic from a burned out testicular *** diagnosis is critical as appropriate therapy can be highly *** present the case of an isolated extragonadal germ cell tumor in the retroperitoneum diagnosed by endoscopic ultrasound-guided fi ne needle *** case underscores the importance of considering germ cell tumors in the differential diagnosis of an unexplained retroperitoneal mass,particularly since immunophenotypic staining may be necessary to establish the diagnosis.2010 *** rights reserved.

关键词： germ cell tumor Endoscopic ultrasound Fine needle aspiration

ASSOCIATION-DNA, A NEW DISCOVERY RELATED TO MALE germ UNIT IN THE POLLEN TUBE OF Camellia sinensis

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Chinese Science Bulletin 1991年第2期36卷 163-166页

作者：莫永胜张玉麟过全生 Department of Biology Wuhan University Wuhan 430072 PRC Division of Basic Courses Zhejiang Agricultural University Hangzhou 310029 PRC

Up to now, male germ units (MGUs)have been found in many angiosperms. This is an important advance in the studies of reproductive biology. However, there is little information on the interaction (the penetration and exchange of substances) between vegetative nucleus (VN) and generative cell (GC) or sperm cells within MGUs. If there exists, what kind of substance Will it be and how will it work, and what physiological function will it have? This is a very important problem which needs to be solved as soon as possible.

关键词： male germ unit association-DNA pollen tube.

Reprogramming of the pig primordial germ cells into pluripotent stem cells: a brief review

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Frontiers of Agricultural Science and Engineering 2019年第1期6卷 28-32页

作者： Qijing LEI Qin PAN Shuai YU Na LI Shulin CHEN Kuldip SIDHU Jinlian HUA College of Veterinary Medicine/Shaanxi Centre of Stem Cells Engineering and Technology Northwest A&F University Centre for Healthy Brain Ageing UNSW Medicine

Primordial germ cells(PGCs) are regarded as unipotent cells that can produce only either spermatogonia or oocytes. However, PGCs can be converted into the pluripotent state by ?rst dedifferentiation to embryonic germ cells and then by reprogramming to induce them to become pluripotent stem cells(iPSCs). These two stages can be completely implemented with mouse cells. However, authentic porcine iPSCs have not been ***, we discuss recent advances in the stem cell ?eld for obtaining iPSCs from PGCs. This knowledge will provide some clues which will contribute to the regulation of reprogramming to pluripotency in farm species.

关键词： pig pluripotent stem cells primordial germ cells reprogramming