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En masse lentiviral gene delivery to mouse fertilized eggs via laser perforation of zona pellucida.


ABSTRACT: Lentiviruses are highly efficient vehicles for delivering genes into cells. They readily transduce primary and immortalized cells in vivo and in vitro. Genes delivered by lentiviruses are incorporated and replicated as part of their host genome and therefore offer a powerful tool for creation of stable cell lines and transgenic animals. However, the zona pellucida surrounding the fertilized eggs acts as a barrier and hinders lentiviral transduction of embryos. Here, we utilize a laser, typically used to perforate the zona pellucida for in vitro fertilization, to permeabilize the zona for lentiviral gene delivery. A single hole in the zona is sufficient for the lentivirus to gain access to fertilized eggs without the need for microinjection for en masse gene delivery. Embryos generated by this method elicit no damage and can develop to term for creation of transgenic animals.

SUBMITTER: Martin NP 

PROVIDER: S-EPMC5990369 | biostudies-literature | 2018 Feb

REPOSITORIES: biostudies-literature

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En masse lentiviral gene delivery to mouse fertilized eggs via laser perforation of zona pellucida.

Martin Negin P NP   Myers Page P   Goulding Eugenia E   Chen Shih-Heng SH   Walker Mitzie M   Porter Thomas M TM   Van Gorder Lucas L   Mathew Amanda A   Gruzdev Artiom A   Romeo Charles C  

Transgenic research 20180213 1


Lentiviruses are highly efficient vehicles for delivering genes into cells. They readily transduce primary and immortalized cells in vivo and in vitro. Genes delivered by lentiviruses are incorporated and replicated as part of their host genome and therefore offer a powerful tool for creation of stable cell lines and transgenic animals. However, the zona pellucida surrounding the fertilized eggs acts as a barrier and hinders lentiviral transduction of embryos. Here, we utilize a laser, typically  ...[more]

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