Transgenic Research

, Volume 4, Issue 4, pp 264–269 | Cite as

An insertional mutation in theBTF3 transcription factor gene leads to an early postimplantation lethality in mice

  • Jian Min Deng
  • Richard R. Behringer


The gene that encodes the general transcription factor known asBTF3 was disrupted in mouse embryonic stem cells in a random mutagenesis screen for developmental genes with the ROSA β-geo (Friedrich and Soriano, 1991) retroviral gene trap vector. TheBTF3 mutation was transmitted through the germline of chimaeric mice. While the endogenousBTF3 gene is ubiquitously expressed, the expression pattern of the β-galactosidase reporter gene present in the gene trap vector inBTF3 heterozygotes was restricted. Mice homozygous for the mutant allele died soon after implantation, around embryonic day 6.5. Thus,BTF3 is essential for postimplantation development. The isolation of theBTF3 sequences in this ROSA β-geo insertion was facilitated by a relatively simple singlelacZ primer reverse transcription PCR strategy.


retrovirus gene traps embryonic stem cells mutagenesis BTF3 


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  1. Behringer, R.R., Crotty, D.A., Tennyson, V.M., Brinster, R.L., Palmiter, R.D. and Wolgemuth, D.J. (1993) Sequences 5′ of the homeobox of theHox-1.4 gene direct tissue-specific expression oflacZ during mouse development.Development 117, 823–33.PubMedGoogle Scholar
  2. Bradley, A., Evans, M., Kaufman, M.H., and Robertson, E. (1984) Formation of germline chimaeras from embryoderived teratocarcinoma cell lines.Nature 309, 255–256CrossRefPubMedGoogle Scholar
  3. Bradley, A. (1987) Production and analysis of chimeric mice. In Robertson, E.J., ed.,Teratocarcinomas and Embryonic Stem Cells: a Practical Approach, pp. 113–151, Oxford: IRL Press.Google Scholar
  4. Capecchi, M.R. (1989) The new mouse genetics: altering the genone by gene targeting.Trends Genet. 5, 70–6.CrossRefPubMedGoogle Scholar
  5. Chirgwin, J.M., Przybyla, A.E., MacDonald, R.J. and Rutter, W.J. (1979) Isolation of biologically active ribonucleic acid from sources enriched in ribonuclease.Biochemistry 18, 5284–99.CrossRefGoogle Scholar
  6. Friedrich, G. and Soriano, P. (1991) Promoter traps in embryonic stem cells: a genetic screen to identify and mutate developmental genes in mice.Genes Dev. 5, 1513–23.PubMedGoogle Scholar
  7. Friedrich, G. and Soriano, P. (1993) Insertional mutagenesis by retroviruses and promoter traps in embryonic stem cells.Methods Enzymol. 225, 681–701.PubMedGoogle Scholar
  8. Gerard, M., Fischer, L., Moncollin, V., Chipoulet, J.M., Chambon, P. and Egly, J.M. (1991) Purification and interaction properties of the human RNA polymerase B(II) general transcription factor BTF2.J. Biol. Chem. 266, 20940–5.PubMedGoogle Scholar
  9. Gossler, A., Joyner, A.L., Rossant, J. and Skarnes, W.C. (1989) Mouse embryonic stem cells and reporter constructs to detect developmentally regulated genes.Science 244, 463–5.PubMedGoogle Scholar
  10. Kanno, M., Chalut, C. and Egly, J.M. (1992) Genomic structure of the putativeBTF3 transcription factor.Gene 117, 219–28.CrossRefPubMedGoogle Scholar
  11. McMahon, A.P. and Bradley, A. (1990). TheWnt-1 (int-1) proto-oncogene is required for development of a large region of the mouse brain.Cell 62, 1073–1085.CrossRefPubMedGoogle Scholar
  12. Parthun, M.R., Mangus, D.A. and Jaehning, J.A. (1992) TheEGD1 product, a yeast homolog of humanBTF3, may be involved in GAL4 DNA binding,Mol. Cell. Biol. 12, 5683–9.PubMedGoogle Scholar
  13. Ronne, H. and Hu, G.Z. (1994) Yeast PBT3 protein is encoded by duplicated genes and inhibits the expression of some genesin vivo.Nucl. Acids Res. 22, 2740–3.PubMedGoogle Scholar
  14. Sambrook, J., Fritsch, E.F. and Maniatis, T. (1989) Molecular cloning: a laboratory manual. Cold Spring Harbor, New York: Cold Spring Harbor Laboratory Press.Google Scholar
  15. Skarnes, W.C., Auerbach, B.A. and Joyner, A. (1992) A gene trap approaching mouse embryonic stem cells: thelacZ reporter is activated by splicing reflects endogenous gene expression, and is mutagenic in mice.Genes Dev. 6, 903–18.PubMedGoogle Scholar
  16. Melcher, H. von, DeGregori, J.V., Rayburn, H., Reddy, S., Friedel, C. and Ruley, H.E. (1992) Selective disruption of genes expressed in totipotent embryonal stem cells.Genes Dev. 6, 919–27.PubMedGoogle Scholar
  17. Zheng, X.M., Moncollin, V., Egly, J.M. and Chambon, P. (1987) A gene transcription factor forms a stable complex with RNA polymerase (II).Cell 50, 361–8.CrossRefPubMedGoogle Scholar
  18. Zheng, X.M., Black, D., Chambon, P. and Egly, J.M. (1990) Sequencing and expression of complementary DNA for the general transcription factorBTF3.Nature 344, 556–9.CrossRefPubMedGoogle Scholar

Copyright information

© Chapman & Hall 1995

Authors and Affiliations

  • Jian Min Deng
    • 1
  • Richard R. Behringer
    • 1
  1. 1.Department of Molecular GeneticsUniversity of Texas, M. D. Anderson Cancer CenterHoustonUSA

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