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

Ivana Gessara, Falk Dittrich, Moritz Hertel, Staffan Hildebrand, Alexander Pfeifer, Carolina Frankl-Vilches, Mike McGrew, Manfred Gahr

Research output: Contribution to journalArticlepeer-review

Abstract

The ability to genetically manipulate organisms has led to significant insights into 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 bloodstream 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. Transduced zfPGCs were injected into the host embryo and transgenic songbirds were successfully generated.

Original languageEnglish
JournalStem Cell Reports
Volume16
Early online date3 Mar 2021
DOIs
Publication statusE-pub ahead of print - 3 Mar 2021

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