Highly Efficient Genome Modification of Cultured Primordial Germ Cells with Lentiviral Vectors to Generate Transgenic Songbirds

Highly efficient genome modification of cultured primordial germ cells with lentiviral vectors to generate transgenic songbirds

10.1101/2020.08.11.245753 ◽

2020 ◽

Author(s):

Ivana Gessara ◽

Falk Dittrich ◽

Moritz Hertel ◽

Staffan Hildebrand ◽

Alexander Pfeifer ◽

...

Keyword(s):

Zebra Finch ◽

Germ Cells ◽

Fluorescent Protein ◽

Vocal Communication ◽

Primordial Germ Cells ◽

Vocal Learning ◽

Lentiviral Vectors ◽

Blood Stream ◽

Stem Cell Markers ◽

Highly Efficient

SUMARYThe ability to genetically manipulate organisms has led to significant insights in functional genomics in many species. In birds, manipulation of the genome is hindered by the inaccessibility of the one-cell embryo. During embryonic development, avian primordial germ cells (PGCs) migrate through the blood stream and reach the gonadal anlage; where they develop into mature germ cells. Here, we explored the use of PGCs to produce transgenic offspring in the zebra finch, which is a major animal model for sexual brain differentiation, vocal learning and vocal communication. Zebra finch PGCs (zfPGCs) obtained from embryonic blood significantly proliferated when cultured in an optimized culture medium and conserved the expression of germ and stem cell markers. Transduction of cultured zfPGCs with lentiviral vectors was highly efficient leading to strong expression of the enhanced green fluorescent protein (eGFP). Transduced zfPGCs were injected into the host embryo and transgenic songbirds were successfully generated.

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Targeting lentiviral vectors to primordial germ cells (PGCs): An efficient strategy for generating transgenic chickens

动物学研究 ◽

10.24272/j.issn.2095-8137.2020.032 ◽

2020 ◽

Vol 41 (3) ◽

pp. 281-291

Author(s):

Zi-Qin Jiang ◽

◽

Han-Yu Wu ◽

Jing Tian ◽

Ning Li ◽

...

Keyword(s):

Germ Cells ◽

Primordial Germ Cells ◽

Lentiviral Vectors ◽

Efficient Strategy ◽

Transgenic Chickens

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

Proceedings, annual meeting, Electron Microscopy Society of America ◽

10.1017/s042482010012374x ◽

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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