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The electronic form of this document may be cited in the following style:
Human Genome Program, U.S. Department of Energy, DOE Human Genome Program Contractor-Grantee Workshop IV, 1994.
Abstracts scanned from text submitted for November 1994 DOE Human Genome Program Contractor-Grantee Workshop. Inaccuracies have not been corrected.
Mouse RNAs for the Determination of Stage- and Tissue-Specific Expression Profiles for Human Genes Throughout Development
Dabney K. Johnson and Lisa J. Stubbs
Mammalian Genetics and Development Section, Biology Division, Oak Ridge National Laboratory, Oak Ridge, Tennessee, 37830
Easy access to tissues for the isolation of mRNA at all developmental stages makes the mouse an ideal model organism for determining the expression profile of a gene throughout the life of a mammal. For cell populations, like those of preimplantation-stage embryos, that are prohibitively difficult to obtain in usable quantities, cRNAs generated from stage-specific cDNA libraries (1) can fill the same role.
We propose, first, to make a series of polyA+ Northern blots that represent the spectrum of mouse development from the fertilized egg to a complete panel of adult tissues. Egg, 2-cell, 8-cell, and blastocyst cRNAs will be generated via in vitro transcription from existing cDNA libraries, while postimplantation mRNAs will be isolated from whole or dissected fetuses, neonates, juveniles, and adults. Given the extensive mouse-human sequence homologies found in most coding sequences, these Northern blots should allow the determination of the expression profile for most human genes in a comparable mammalian system. In addition, aliquots of these same stage-specific RNAs will be preserved for use in RT-PCR reactions in cases where gene-specific primers can be used.
(1) Rothstein, J.L., Johnson, D.K., DeLoia, J.A., Skowronski, J., Solter, D., and Knowles, B.B. (1992) Gene expression during preimplantation mouse development. Genes and Devel. 6, 1190-1201.