cDNA Libraries from Single Human Preimplantation Embryos

Adjaye, James; Daniels, Rob; Bolton, Virginia N.; Monk, Marilyn

doi:10.1006/geno.1997.5117

Cited by 41 publications

(11 citation statements)

References 46 publications

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“…To illustrate this point, of potential significance is that of the 50 000 to 100 000 genes in each cell, up to 30 000 are expected to be expressed in the brain (Adams et al, 1991). A large proportion of embryo transcripts (14·4 per cent) have already been found to be homologous to brain transcripts (Adams et al, 1991), their expression initiated in the preimplantation embryo (Adjaye et al, 1997). The technical challenge will be to dissect the genetic structuring and functioning of the brain; where better to begin than the human preimplantation embryo.…”

Section: Discussionmentioning

confidence: 99%

“…However, the criticism that recombinant DNA technology may not be sufficiently sensitive to generate definitive molecular information from single human preimplantation embryos is continually being challenged. The ability to construct stagespecific cDNA libraries provides an important new resource for the investigation of the molecular programming of early human development (Belyvasky et al, 1989;Buraczynska et al, 1995;Adjaye et al, 1997Adjaye et al, , 1998. Such stage-specific embryo libraries display between 100 000 and a million clones, sufficient to represent the entire active gene population at these early stages of human development and would highlight the use of the human embryo itself for studies of specific differentiation studies.…”

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confidence: 99%

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The expression of genes in human preimplantation embryos

Pergament

Fiddler

1998

Prenat. Diagn.

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The study of gene expression in human preimplantation embryos is establishing itself as a necessary dimension of developmental biology and medical genetics. Transcripts identified in human preimplantation embryos include housekeeping genes, transcription and growth factor genes, sex‐determining genes, tissue‐specific genes and novel genes, as well as genes of unknown function. Strategies are being developed which will eventually permit the most sophisticated gene expression studies on single human embryos of co‐ordinated transcription and translational regulation. There is both a need for international co‐operation for the systematic construction of expression maps and a need to establish databases of expression patterns during different stages of human development. Understanding how genes are regulated in humans is essential for understanding both normal development and disease. Until recently, studies of gene expression and regulation during embryogenesis were almost exclusively limited to prokaryotes and to eukaryotes other than man. The introduction of artificial reproductive technologies in conjunction with the development of recombinant molecular technologies applicable to single cells has made possible the study of human development at its earliest stages (Pergament and Bonnicksen, 1994). Although there are still enormous technical challenges, robust strategies have been, and continue to be, developed for connecting DNA sequence to such endophenotypes as timing and level of gene expression at the single cell level. Questions currently being asked in human developmental genetic studies concern the pronucleus, the zygote and the preimplantation embryo: what genes are expressed? When are they expressed? What functions do they perform and how, in sequence or in combination? And, what elements control and regulate their expression? This review provides an overview of current knowledge about the expression of different embryonic genes during early human development and discusses future prospects, which includes a need for international co‐operation similar to the Human Genome Project. Copyright © 1998 John Wiley & Sons, Ltd.

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Section: Discussionmentioning

confidence: 99%

mentioning

confidence: 99%

The expression of genes in human preimplantation embryos

Pergament

Fiddler

1998

Prenat. Diagn.

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“…Protocols used for sample preparation, reverse transcriptase (RT) and PCR amplification have been described previously by Liang & Pardee 1992, Adjaye et al . 1997, Daniels et al .…”

Section: Methodsmentioning

confidence: 99%

Analysis of gene expression in rabbit nuclear transfer embryos: Use of single‐embryo mRNA differential display

et al. 2003

Dev Growth Differ

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Lack of or abnormal expression of developmentally important genes is believed to hamper early development of the nuclear transfer (NT) embryo. To identify stage-specific genes in rabbit NT embryo development, mRNA differential display was used to compare the mRNA content of rabbit NT embryos at different developmental stages, from Metaphase II oocytes to 8-16-cell stage embryos. Thirty-four zygotic transcripts, which abruptly appeared at the 8-16-cell stage in rabbit NT embryos, were isolated; 11 of these were potential novel genes with no matches in the current databases. Of the remaining 23, 12 were matched with established sequence tags with functions uncharacterized and the other 11 were homologous to those in the European Molecular Biology Laboratory (EMBL) and GenBank databases . The differential expression of eight of the 34 amplicons were confirmed by reverse Northern blotting, and four positive clones were validated. Previous studies and present data indicated that these three genes were probably related to preimplantation rabbit embryo development.

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“…The first cDNA library of single human preimplantation embryos for transcriptome analysis was published in 1997, 1999 . A targeted profiling for known transcripts was then accessible using quantitative PCR or microarray approaches.…”

Section: Growing Heterogeneity In the Early Preimplantation Embryomentioning

confidence: 99%

New insights into human primordial germ cells and early embryonic development from single‐cell analysis

2017

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Human preimplantation developmental studies are difficult to accomplish due to associated ethical and moral issues. Preimplantation cells are rare and exist only in transient cell states. From a single cell, it is very challenging to analyse the origination of the heterogeneity and complexity inherent to the human body. However, recent advances in single-cell technology and data analysis have provided new insights into the process of early human development and germ cell specification. In this Review, we examine the latest single-cell datasets of human preimplantation embryos and germ cell development, compare them to bulk cell analyses, and interpret their biological implications.

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cDNA Libraries from Single Human Preimplantation Embryos

Cited by 41 publications

References 46 publications

The expression of genes in human preimplantation embryos

The expression of genes in human preimplantation embryos

Analysis of gene expression in rabbit nuclear transfer embryos: Use of single‐embryo mRNA differential display

New insights into human primordial germ cells and early embryonic development from single‐cell analysis

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