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Laser-assisted Lentiviral Gene Delivery to Mouse Fertilized Eggs
Published on: November 1, 2018
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AAV diffuses across zona pellucida for effortless gene delivery to fertilized eggs
Charles Romeo1, Shih-Heng Chen1, Eugenia Goulding1
1Neurobiology Laboratory, Viral Vector Core, National Institute of Environmental Health Sciences, NIH/DHHS, 111 T.W. Alexander Drive, Research Triangle Park, N.C, 27709, USA.
Biochemical and Biophysical Research Communications
|March 22, 2020
Summary
Adeno-associated viruses (AAVs) can efficiently deliver genes into fertilized mammalian eggs by diffusing through the zona pellucida. This method bypasses costly micromanipulation techniques, though embryo development rates may be improved by later-stage exposure.
Area of Science:
- Developmental Biology
- Gene Therapy
- Animal Biotechnology
Background:
- Gene delivery to fertilized eggs is crucial for creating transgenic animals and studying early development.
- The zona pellucida presents a significant barrier to efficient gene delivery, necessitating complex and expensive methods.
- Existing techniques like micromanipulation, laser perforation, and electroporation require specialized equipment and expertise.
Purpose of the Study:
- To investigate the potential of adeno-associated viruses (AAVs) for efficient gene delivery to mammalian fertilized eggs.
- To determine if AAVs can penetrate the zona pellucida without invasive manipulation of the eggs.
- To assess the impact of AAV gene delivery on early embryo development and identify methods to mitigate adverse effects.
Main Methods:
- Fertilized mammalian eggs were exposed to adeno-associated virus (AAV) serotypes 1 and DJ.
- Gene delivery efficiency across the zona pellucida was assessed without any micromanipulation.
- Embryo development rates were monitored after AAV treatment at different stages of in vitro development.
Main Results:
- AAV serotypes 1 and DJ demonstrated efficient diffusion across the zona pellucida for gene delivery.
- All tested AAV serotypes led to reduced embryo development rates.
- Exposing embryos to AAVs at later stages of in vitro development mitigated adverse effects on embryo development.
Conclusions:
- Adeno-associated viruses (AAVs) offer a non-invasive method for gene delivery to fertilized mammalian eggs.
- AAVs can bypass the zona pellucida barrier, simplifying gene delivery protocols.
- AAVs are a promising tool for transient gene delivery due to their low immunogenicity and lack of chromosomal integration.
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