Mycoplasma-mediated uptake of the exogenous human BRCA1 gene by hatching blastocysts

Purpose: Biological vectors for cell transfection are mainly viral in origi n, with inherent shortcomings. Mycoplasmas are ubiquitous organisms that tr averse cells easily The objective was to determine if Ureaplasma urealyticu m (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 T he blastocysts were exposed to human BRCA1 DNA lacking homology in the mous e genome for 2 hr, followed by DNase-I treatment and wash. Polymerase chain reaction and agarose gel electrophoresis analysis of amplified products we re performed. Results: The BRCA1 gene was detected in the blastocysts only when viable Ur eaplasma was present. PCR analyses of control Ureaplasma and untreated blas tocysts were negative. Conclusion: Viable Ureaplasma organisms were shown to mediate the uptake of DNA fragments into blastocysts resulting in transgenic mouse blastocysts w ith a normal human BRCA1 exon II gene.