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Mycoplasma-mediated uptake of the exogenous human BRCA1 gene by hatching blastocysts
P J Chan1, J E Brossfield, W C Patton
1Department of Gynecology and Obstetrics, Loma Linda University School of Medicine, California 92350, USA.
Purpose:
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.
Conclusion:
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.