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 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.
Reproductive Genetics Institute, Illinois Masonic Medical Center, Chicago 60657, USA.
Y
Y
Verlinsky
G
G
Morozov
O
O
Verlinsky
V
V
Koukharenko
S
S
Rechitsky
E
E
Goltsman
V
V
Ivakhnenko
V
V
Gindilis
C M
CM
Strom
A
A
Kuliev
D001755:Blastocyst / Q000737:chemistry*; D003001:Cloning, Molecular; D018076:DNA, Complementary / Q000096:biosynthesis / Q000235:genetics*; D005314:Embryonic and Fetal Development / Q000235:genetics; D018507:Gene Expression Regulation, Developmental; D015723:Gene Library; D006801:Humans; D009024:Morphogenesis / Q000235:genetics; D016133:Polymerase Chain Reaction; D012333:RNA, Messenger / Q000302:isolation & purification*
D018076:DNA, Complementary; D012333:RNA, Messenger