No abstract

25 June 2012.

Retrovirus vectors offer a simple and highly efficient method for introducing new genes into mammalian cells. Here, we have examined the efficiency of gene transfer into hematopoietic cells with retrovirus vectors carrying the neomycin (neo) resistance gene expressed from different transcriptional regulatory regions. Direct infection of mouse bone marrow cells resulted in high efficiencies of gene transfer into a variety of myeloid progenitor cells, including pluripotent, erythroid, and granulocyte-macrophage colony-forming cells with all the vectors examined. However, the progeny derived from individual pluripotent progenitor cells infected with different vectors differed markedly in the proportion of G418-resistant progenitor cells, as judged by their ability to survive selection in the drug G418. This biological assay suggests that the highest level of expression was observed when the neo gene was expressed from constructs that contained the herpes thymidine kinase promoter rather than the viral long terminal repeat or the simian virus 40 early region promoter. In contrast, neo gene expression was highest in fibroblasts infected with the vector containing the simian virus 40 early region promoter. These results show that high and sustainable levels of gene expression in hematopoietic cells can be obtained with retrovirus vectors containing appropriate transcriptional regulatory regions.

(Abstracted from Human Reproduction 2018;33(9):1767–1776) Preimplantation genetics for aneuploidy (PGT-A) has shown mixed results on pregnancy rates when applied at the cleavage stage of development due to a high rate of mosaicism and limitations of the comprehensive chromosomal screening technology, FISH, which was available at the time. A pilot study aiming to address this found that array comparative genomic hybridization (aCGH) analysis of both polar bodies possessed 94% concordance with the chromosome analysis of the resulting fertilized oocytes.