Isolation of cDNA libraries from individual human preimplantation embryos

Although available data from the mouse model suggest that morphogenesis during preimplantation development is dependent on the expression of embryonic genes, convincing data for human preimplantation embryos are missing. To investigate the expression of genes involved in human preimplantation develo...

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Bibliographic Details
Published in:Molecular human reproduction 1998-06, Vol.4 (6), p.571-575
Main Authors: Verlinsky, Y, Morozov, G, Verlinsky, O, Koukharenko, V, Rechitsky, S, Goltsman, E, Ivakhnenko, V, Gindilis, V, Strom, C M, Kuliev, A
Format: Article
Language:English
Subjects:
Blastocyst - chemistry
Cloning, Molecular
DNA, Complementary - biosynthesis
DNA, Complementary - genetics
Embryonic and Fetal Development - genetics
Gene Expression Regulation, Developmental
Gene Library
Humans
Morphogenesis - genetics
Polymerase Chain Reaction
RNA, Messenger - isolation & purification
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Summary:Although available data from the mouse model suggest that morphogenesis during preimplantation development is dependent on the expression of embryonic genes, convincing data for human preimplantation embryos are missing. To investigate the expression of genes involved in human preimplantation development we constructed cDNA libraries from human individual blastocysts and screened them for the expression of beta-actin, CD59, homeoboxes OCT-3 and HOXA4, and HLA-G and hMLH-1 genes. Beta-actin, CD59, and OCT-3 were detected by reverse transcription-polymerase chain reaction (RT-PCR), while HOXA4, HLA-G and hMLH-1 were undetected. Sequencing of 48 random clones from two libraries revealed a different identity to the known genes including 99% identity to human histone 3.1 and human ribosome protein S25 complete cDNA. These data demonstrate the feasibility of constructing cDNA libraries from individual human preimplantation embryos and their potential usefulness in the assessment of the relevance of specific gene expression in the failures of preimplantation development.
ISSN:1360-9947
1460-2407
DOI:10.1093/molehr/4.6.571