Production of Transgenic Chickens Using Cultured Primordial Germ Cells and Gonocytes.

Production of Transgenic Chickens Using Cultured Primordial Germ Cells and Gonocytes.
Methods Mol Biol. 2019;1874:403-430
Authors: Collarini EJ, Leighton PA, Van de Lavoir MC
Abstract
The unique characteristics of the avian embryo, with its large opaque yolk, have necessitated the development of different approaches to transgenesis from those that have been successful in mammalian species. Genetic modification of birds was greatly advanced by the ability to grow long-term cultures of primordial germ cells (PGCs). These cells are obtained from embryos, established in culture, and can be propagated without losing the ability to contribute to the germline when reintroduced into a host animal. PGCs can be genetically modified in culture using traditional transfection and selection techniques, including gene targeting and site-specific nuclease approaches. Here, we describe our methods for deriving cell lines, long-term culture, genetic modification, production of germline chimeras and obtaining fully transgenic birds with the desired genetic modifications.
PMID: 30353528 [PubMed - in process]

Source: Mol Biol Cell - October 27, 2018 Category: Molecular Biology Authors: Collarini EJ, Leighton PA, Van de Lavoir MC Tags: Methods Mol Biol Source Type: research