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Published on: November 5, 2012
Mammalian transgenesis by intracytoplasmic sperm injection
A C Perry1, T Wakayama, H Kishikawa
1Department of Anatomy and Reproductive Biology, University of Hawaii School of Medicine, Honolulu, HI 96822, USA. perry@hawaii.edu
Science (New York, N.Y.)
|May 15, 1999
Summary
Sperm heads can deliver foreign DNA into mouse eggs, creating reporter gene-expressing embryos and offspring. This adaptable method offers a new way to achieve transgenesis efficiently.
Area of Science:
- Reproductive biology
- Molecular genetics
- Developmental biology
Background:
- Transgenesis, the process of introducing foreign DNA into an organism, is crucial for genetic research and biotechnology.
- Existing methods for transgenesis can be inefficient or technically demanding.
- Efficient delivery of exogenous DNA into oocytes is a key challenge.
Purpose of the Study:
- To investigate the feasibility of using sperm heads as carriers for exogenous DNA delivery into unfertilized mouse oocytes.
- To establish a reproducible method for transgenesis using sperm-mediated DNA transfer.
- To assess the efficiency of transgene expression in resulting embryos and offspring.
Main Methods:
- Coinjection of unfertilized mouse oocytes with sperm heads and exogenous DNA encoding reporter genes (GFP or beta-galactosidase).
- Microinjection of sperm heads pre-associated with exogenous DNA into oocytes.
- Nonselective transfer of resulting embryos to surrogate mothers for gestation.
- Analysis of transgene integration and expression in embryos and offspring.
Main Results:
- Coinjection resulted in 64-94% transgene-expressing embryos, indicating efficient DNA-sperm head association and oocyte delivery.
- Approximately 20% of offspring from the GFP series expressed the integrated transgene.
- Reporter gene expression confirmed successful delivery and integration of exogenous DNA.
Conclusions:
- Sperm heads can be effectively utilized to deliver exogenous DNA into oocytes.
- This sperm-mediated DNA transfer method is adaptable and reproducible for transgenesis.
- The technique holds promise for advancing genetic engineering and research applications.
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