Sperm cells as vectors for introducing foreign DNA into eggs: genetic transformation of mice.
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.
Lavitrano
M
M
Institute of Biomedical Technology, CNR, Rome, Italy.
Camaioni
A
A
Fazio
V M
VM
Dolci
S
S
Farace
M G
MG
Spadafora
C
C
D000818:Animals; D001483:Base Sequence; D015500:Chloramphenicol O-Acetyltransferase / Q000235:genetics*; D004247:DNA / Q000235:genetics*; D005260:Female; D005796:Genes; D005798:Genes, Bacterial; D005822:Genetic Vectors; D008297:Male; D051379:Mice; D008822:Mice, Transgenic; D008969:Molecular Sequence Data; D009928:Organ Specificity; D010063:Ovum / Q000502:physiology*; D013084:Sperm-Ovum Interactions; D013094:Spermatozoa / Q000502:physiology*
D004247:DNA; D015500:Chloramphenicol O-Acetyltransferase