In vivo transfection of testicular germ cells and transgenesis by using the mitochondrially localized jellyfish fluorescent protein gene.
We aimed to introduce foreign DNA into spermatogenic cells in the testis by injection of the DNA encoding jellyfish fluorescent proteins, green fluorescent protein (GFP) and yellow fluorescent protein (YFP) into the seminiferous tubules and in vivo electroporation. We obtained fluorescent spermatozoa only when using the gene of the YFP protein fused to a mitochondrial localization signal peptide. Intracytoplasmic injection into oocytes of these spermatozoa gave fluorescent fetuses and pups. Almost all of the individuals produced from fluorescent spermatozoa were transgenic. We confirmed integration of the gene into chromosomes and its transmission into offspring. This is the first report of gene transfer into germ cells and subsequent production of transgenic offspring.
Huang
Z
Z
Mammalian Development Laboratory, National Institute of Genetics, Mishima, Japan.
Tamura
M
M
Sakurai
T
T
Chuma
S
S
Saito
T
T
Nakatsuji
N
N
D000818:Animals; D001426:Bacterial Proteins / Q000032:analysis / Q000235:genetics; D018274:Electroporation; D018014:Gene Transfer Techniques; D017930:Genes, Reporter; D049452:Green Fluorescent Proteins; D008164:Luminescent Proteins / Q000032:analysis / Q000235:genetics*; D008297:Male; D051379:Mice; D008810:Mice, Inbred C57BL; D008811:Mice, Inbred DBA; D008813:Mice, Inbred ICR; D008928:Mitochondria / Q000502:physiology* / Q000648:ultrastructure; D021382:Protein Sorting Signals; D037401:Scyphozoa; D012671:Seminiferous Tubules / Q000502:physiology*; D020554:Sperm Injections, Intracytoplasmic; D013094:Spermatozoa / Q000502:physiology*
D001426:Bacterial Proteins; D008164:Luminescent Proteins; D021382:Protein Sorting Signals; C054562:yellow fluorescent protein, Bacteria; D049452:Green Fluorescent Proteins