Germline transmission of genetically modified primordial germ cells

Nature ◽  
2006 ◽  
Vol 441 (7094) ◽  
pp. 766-769 ◽  
Author(s):  
Marie-Cecile van de Lavoir ◽  
Jennifer H. Diamond ◽  
Philip A. Leighton ◽  
Christine Mather-Love ◽  
Babette S. Heyer ◽  
...  
Keyword(s):  
Germ Cells ◽  
Genetically Modified ◽  
Primordial Germ Cells ◽  
Germline Transmission

scholarly journals Interspecific Germline Transmission of Cultured Primordial Germ Cells

PLoS ONE ◽  
2012 ◽  
Vol 7 (5) ◽  
pp. e35664 ◽  
Author(s):  
Marie-Cecile van de Lavoir ◽  
Ellen J. Collarini ◽  
Philip A. Leighton ◽  
Jeffrey Fesler ◽  
Daniel R. Lu ◽  
...  
Keyword(s):  
Germ Cells ◽  
Primordial Germ Cells ◽  
Germline Transmission

scholarly journals Improved Germline Transmission in Chicken Chimeras Produced by Transplantation of Gonadal Primordial Germ Cells into Recipient Embryos1

2003 ◽  
Vol 68 (5) ◽  
pp. 1657-1662 ◽  
Author(s):  
Tae Sub Park ◽  
Dong Kee Jeong ◽  
Jin Nam Kim ◽  
Gwon Hwa Song ◽  
Yeong Ho Hong ◽  
...  
Keyword(s):  
Germ Cells ◽  
Primordial Germ Cells ◽  
Germline Transmission

scholarly journals Characterisation and Germline Transmission of Cultured Avian Primordial Germ Cells

PLoS ONE ◽  
2010 ◽  
Vol 5 (11) ◽  
pp. e15518 ◽  
Author(s):  
Joni Macdonald ◽  
James D. Glover ◽  
Lorna Taylor ◽  
Helen M. Sang ◽  
Michael J. McGrew
Keyword(s):  
Germ Cells ◽  
Primordial Germ Cells ◽  
Germline Transmission

scholarly journals Migration and Proliferation of Intact and Genetically Modified Primordial Germ Cells and the Generation of a Transgenic Chicken1

2010 ◽  
Vol 82 (2) ◽  
pp. 257-262 ◽  
Author(s):  
Jin Nam Kim ◽  
Tae Sub Park ◽  
Sang Hyun Park ◽  
Kyung Je Park ◽  
Tae Min Kim ◽  
...  
Keyword(s):  
Germ Cells ◽  
Genetically Modified ◽  
Primordial Germ Cells

scholarly journals The reversible developmental unipotency of germ cells in chicken

Reproduction ◽  
2010 ◽  
Vol 139 (1) ◽  
pp. 113-119 ◽  
Author(s):  
Jin Gyoung Jung ◽  
Young Mok Lee ◽  
Jin Nam Kim ◽  
Tae Min Kim ◽  
Ji Hye Shin ◽  
...  
Keyword(s):  
Stem Cells ◽  
Germ Cell ◽  
Germ Cells ◽  
Developmental Stages ◽  
Production Systems ◽  
Primordial Germ Cells ◽  
Transmission Efficiency ◽  
Germline Transmission ◽  
Adult Testis

We recently developed bimodal germline chimera production approaches by transfer of primordial germ cells (PGCs) or embryonic germ cells (EGCs) into embryos and by transplantation of spermatogonial stem cells (SSCs) or germline stem cells (GSCs) into adult testes. This study was undertaken to investigate the reversible developmental unipotency of chicken germ cells using our established germline chimera production systems. First, we transferred freshly isolated SSCs from adult testis or in vitro cultured GSCs into stage X and stage 14–16 embryos, and we found that these transferred SSCs/GSCs could migrate to the recipient embryonic gonads. Of the 527 embryos that received SSCs or GSCs, 135 yielded hatchlings. Of 17 sexually mature males (35.3%), six were confirmed as germline chimeras through testcross analysis resulting in an average germline transmission efficiency of 1.3%. Second, PGCs/EGCs, germ cells isolated from embryonic gonads were transplanted into adult testes. The EGC transplantation induced germline transmission, whereas the PGC transplantation did not. The germline transmission efficiency was 12.5 fold higher (16.3 vs 1.3%) in EGC transplantation into testis (EGCs to adult testis) than that in SSC/GSC transfer into embryos (testicular germ cells to embryo stage). In conclusion, chicken germ cells from different developmental stages can (de)differentiate into gametes even after the germ cell developmental clock is set back or ahead. Use of germ cell reversible unipotency might improve the efficiency of germ cell-mediated germline transmission.

scholarly journals X-irradiation Removes Endogenous Primordial Germ Cells (PGCs) and Increases Germline Transmission of Donor PGCs in Chimeric Chickens

2012 ◽  
Vol 58 (4) ◽  
pp. 432-437 ◽  
Author(s):  
Yoshiaki NAKAMURA ◽  
Fumitake USUI ◽  
Daichi MIYAHARA ◽  
Takafumi MORI ◽  
Tamao ONO ◽  
...  
Keyword(s):  
Germ Cells ◽  
Primordial Germ Cells ◽  
Germline Transmission ◽  
X Irradiation

scholarly journals Successful CRISPR/Cas9 mediated homologous recombination in a chicken cell line

F1000Research ◽  
2018 ◽  
Vol 7 ◽  
pp. 238 ◽  
Author(s):  
Ekaterina Antonova ◽  
Olga Glazova ◽  
Anna Gaponova ◽  
Aykaz Eremyan ◽  
Svetlana Zvereva ◽  
...  
Keyword(s):  
Gene Expression ◽  
Germ Cells ◽  
Genetically Modified ◽  
Primordial Germ Cells ◽  
Fluorescent Microscopy ◽  
Constitutive Expression ◽  
Pcr Analysis ◽  
Tissue Specific ◽  
Knock Out ◽  
Chicken Cell

Background: CRISPR/Cas9 system is becoming the dominant genome editing tool in a variety of organisms. CRISPR/Cas9 mediated knock out has been demonstrated both in chicken cell lines and in chicken germ cells that served to generate genetically modified birds. However, there is limited data about CRISPR/Cas9 dependent homology directed repair (HDR) for avian, even in cell culture. Few attempts have been made with integrations in safe harbor loci of chicken genome that induces constitutive expression of the inserted gene. Gene expression under an endogenous promoter would be more valuable than under a constitutive exogenous promoter, as it allows the gene expression to be tissue-specific. Methods: Three gRNAs were chosen to target chicken 3’-untranslated region of GAPDH gene. Cas9-mediated activity in the targeted locus for the gRNAs in DF-1 cells was estimated by T7E1 assay. To edit the locus, the HDR cassette was added along with CRISPR/Cas9. The inserted sequence contained eGFP in frame with a GAPDH coding sequence via P2A and Neomycin resistance gene (neoR) under cytomegalovirus promoter. Correct integration of the cassette was confirmed with fluorescent microscopy, PCR analysis and sequencing. Enrichment of modified cells was done by G418 selection. Efficiency of integration was assessed with fluorescence activated cell sorting (FACS). Results: We have established a CRISPR/Cas9 system to target an endogenous locus and precisely insert a gene under endogenous control. In our system, we used positive and negative selection to enrich modified cells and remove cells with undesirable insertions. The efficiency of CRISPR/Cas9-mediated HDR was increased up to 90% via G418 enrichment. We have successfully inserted eGFP under control of the chicken GAPDH promoter. Conclusions: The approach can be used further to insert genes of interest under control of tissue-specific promoters in primordial germ cells in order to produce genetically modified birds with useful for biotechnological purposes features.

scholarly journals Successful CRISPR/Cas9 mediated homologous recombination in a chicken cell line

F1000Research ◽  
2018 ◽  
Vol 7 ◽  
pp. 238 ◽  
Author(s):  
Ekaterina Antonova ◽  
Olga Glazova ◽  
Anna Gaponova ◽  
Aykaz Eremyan ◽  
Svetlana Zvereva ◽  
...  
Keyword(s):  
Gene Expression ◽  
Germ Cells ◽  
Genetically Modified ◽  
Primordial Germ Cells ◽  
Fluorescent Microscopy ◽  
Constitutive Expression ◽  
Pcr Analysis ◽  
Tissue Specific ◽  
Knock Out ◽  
Chicken Cell

Background: CRISPR/Cas9 system is becoming the dominant genome editing tool in a variety of organisms. CRISPR/Cas9 mediated knock out has been demonstrated both in chicken cell lines and in chicken germ cells that served to generate genetically modified birds. However, there is limited data about CRISPR/Cas9 dependent homology directed repair (HDR) for avian, even in cell culture. Few attempts have been made with integrations in safe harbor loci of chicken genome that induces constitutive expression of the inserted gene. Gene expression under an endogenous promoter would be more valuable than under a constitutive exogenous promoter, as it allows the gene expression to be tissue-specific. Methods: Three gRNAs were chosen to target chicken 3’-untranslated region of GAPDH gene. Cas9-mediated activity in the targeted locus for the gRNAs in DF-1 cells was estimated by T7E1 assay. To edit the locus, the HDR cassette was added along with CRISPR/Cas9. The inserted sequence contained eGFP in frame with a GAPDH coding sequence via P2A and Neomycin resistance gene (neoR) under cytomegalovirus promoter. Correct integration of the cassette was confirmed with fluorescent microscopy, PCR analysis and sequencing. Enrichment of modified cells was done by G418 selection. Efficiency of integration was assessed with fluorescence activated cell sorting (FACS). Results: We have established a CRISPR/Cas9 system to target an endogenous locus and precisely insert a gene under endogenous control. In our system, we used positive and negative selection to enrich modified cells and remove cells with undesirable insertions. The efficiency of CRISPR/Cas9-mediated HDR was increased up to 90% via G418 enrichment. We have successfully inserted eGFP under control of the chicken GAPDH promoter. Conclusions: The approach can be used further to insert genes of interest under control of tissue-specific promoters in primordial germ cells in order to produce genetically modified birds with useful for biotechnological purposes features.

Hexachlorobenzene toxicity in the rat ovary: II. Ultrastructure induced by medium (10 mg/kg) dose exposure

1992 ◽  
Vol 50 (1) ◽  
pp. 668-669
Author(s):  
Amreek Singh ◽  
Warren G. Foster ◽  
Anna Dykeman ◽  
David C. Villeneuve
Keyword(s):  
Germ Cells ◽  
Primordial Germ Cells ◽  
Surface Epithelium ◽  
Morphological Parameters ◽  
Comprehensive Investigation ◽  
Ovary Surface Epithelium ◽  
Rat Ovary ◽  
High Doses

Hexachlorobenzene (HCB) is a known toxicant that is found in the environment as a by-product during manufacture of certain pesticides. This chlorinated chemical has been isolated from many tissues including ovary. When administered in high doses, HCB causes degeneration of primordial germ cells and ovary surface epithelium in sub-human primates. A purpose of this experiment was to determine a no-effect dose of the chemical on the rat ovary. The study is part of a comprehensive investigation on the effects of the compound on the biochemical, hematological, and morphological parameters in the monkey and rat.

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