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Published on: May 17, 2020
Sperm cells as vectors for introducing foreign DNA into eggs: genetic transformation of mice
M Lavitrano1, A Camaioni, V M Fazio
1Institute of Biomedical Technology, CNR, Rome, Italy.
Abstract:
Mature mouse sperm cells incubated in an isotonic buffer with cloned DNA capture DNA molecules over a 15 min period. Spermatozoa incubated with pSV2CAT plasmid in either circular or linear form were used to fertilize mouse eggs in vitro. Sequences complementary to pSV2CAT were identified in approximately 30% of 250 progeny by Southern blotting. A genomic library was constructed from the DNA of a positive mouse. Three positive clones were identified and two adjacent HincII restriction fragments of 240 and 370 bp showed identical sequences to the corresponding fragments of the pSV2CAT plasmid. F1 progeny showed paternal and maternal transmission of the transgenes from founders. CAT gene expression was detected on tissues of adult F1 individuals, preferentially on tails and muscle. We conclude that transgenic mice can be obtained using sperm cells as foreign DNA vectors.
Insights
Sperm cells can deliver cloned DNA into mouse eggs, creating transgenic mice. This method successfully integrated and expressed foreign DNA, demonstrating a novel approach for genetic modification in mammals.
Area of Science:
- Reproductive Biology
- Molecular Genetics
- Transgenesis
Background:
- Sperm cells possess the ability to bind and internalize exogenous DNA molecules.
- Efficient methods for generating transgenic animals are crucial for biological research and therapeutic applications.
Purpose of the Study:
- To investigate the potential of using mature mouse sperm cells as vectors for foreign DNA delivery.
- To determine if sperm-mediated DNA transfer can result in the generation of viable transgenic mice.
Main Methods:
- Mature mouse sperm were incubated with pSV2CAT plasmid DNA.
- Spermatozoa were used for in vitro fertilization of mouse eggs.
- Southern blotting was employed to detect the presence of foreign DNA in progeny.
- A genomic library was constructed from positive mice for further analysis.
- CAT gene expression was assessed in F1 individuals.
Main Results:
- Sperm cells captured cloned DNA within a 15-minute incubation period.
- Approximately 30% of the resulting progeny (250 mice) contained sequences complementary to the pSV2CAT plasmid.
- Analysis of positive mice revealed integration of the foreign DNA, with identical sequences to the plasmid found in restriction fragments.
- Transgenes were transmitted paternally and maternally to the F1 generation.
- CAT gene expression was detected in adult F1 mice, particularly in tail and muscle tissues.
Conclusions:
- Mature mouse sperm can function as effective vectors for delivering foreign DNA.
- Sperm-mediated gene transfer is a viable method for producing transgenic mice.
- This technique offers a novel approach for genetic engineering in mammals, with potential applications in research and biotechnology.
