Mycoplasma-mediated uptake of the exogenous human BRCA1 gene by hatching blastocysts.
OBJECTIVE
Biological vectors for cell transfection are mainly viral in origin, with inherent shortcomings. Mycoplasmas are ubiquitous organisms that traverse cells easily. The objective was to determine if Ureaplasma urealyticum (T-mycoplasma) would vector exogenous BRCA1 DNA into blastocysts.


METHODS
Hatching mouse blastocysts (N = 70) were incubated in the presence of either viable or dead Ureaplasma urealyticum at 37 degrees C for 1 hr. The blastocysts were exposed to human BRCA1 DNA lacking homology in the mouse genome for 2 hr, followed by DNase-1 treatment and wash. Polymerase chain reaction and agarose gel electrophoresis analysis of amplified products were performed.


RESULTS
The BRCA1 gene was detected in the blastocysts only when viable Ureaplasma was present. PCR analyses of control Ureaplasma and untreated blastocysts were negative.


CONCLUSIONS
Viable Ureaplasma organisms were shown to mediate the uptake of DNA fragments into blastocysts, resulting in transgenic mouse blastocysts with a normal human BRCA1 exon 11 gene.
Chan
P J
PJ
Department of Gynecology and Obstetrics, Loma Linda University School of Medicine, California 92350, USA.
Brossfield
J E
JE
Patton
W C
WC
King
A
A
D000818:Animals; D019313:BRCA1 Protein / Q000235:genetics*; D001755:Blastocyst / Q000166:cytology*; D005822:Genetic Vectors; D006801:Humans; D051379:Mice; D016133:Polymerase Chain Reaction; D014162:Transfection / Q000235:genetics*; D016990:Ureaplasma urealyticum / Q000235:genetics*
D019313:BRCA1 Protein