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In situ freezing of embryonic stem cells in multiwell plates
~]~ECHNICAL unexpected; many h o m e o b o x proteins have similar DNA binding specificities, and ectopic expression suggests large overlap in target genes, The expression patterns of the genes give few indications to their likely ffmction. Genes regulated by homcotic genes are likcly to govern general morphogenetic processes such as mitotic rate, mitotic orientation, cell-cell interactions and cuticular differentiationS. The real power of this approach lies in the combination of biochemical techniques and genetic analysis to assess whether candidate genes isolated are likely to be functional targets in vivo. The method should be generally applicable to the investigation of
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developmental systems in other species, and enable the isolation of binding sites of other types of regulatory proteins. Acknowledgements l.J.J, is a Lister fellow. References 1 Garcia-Bellido, A. (1975) in Cell Patternitzg (Ciba Foundation Syrup. 29), pp. 161-178, Elsevier 2 Scott, MP., Tamkun, J.W. and Hartzell. G.W., III (1989) Biocbim Biophys. Acta 989, 25q8 3 Kissinger, C.R. etal. (1990) Cel163, ~75-590 4 lmmergluck, K., Lawrence, EA. and Bienz, M. (19~)) Cell62, 261-268 5 Reuter, R., Panganiban, G.E.F., Hoffman, F.M. and Scott, M.P.
(1990) Development 110, 1031-1040 6 Pollwein, 1:, Wagner, S. and Knippers, R. (1987) Nucleic Acids Res. 15, 9741-9759 7 Gruss, C. etal. (1988) Virolog)' 167, 349-360 8 Sompayrac, L. and Danna, K.J. (1990) Proc. Nat/A cad. Sci. I,X4 87, 3274-3278 9 Oliphant, A.R., Brandl, C0. and Struhl, K. (1989) Mol. Cell. Biol. 9, 2944-2949 10 Kinzler, K.W. and Vogelstein, B. (1990) Mol. Cell. Biol. 10, 634-642 11 Laughon, AP., Howell, W. and Scott, M.P. (1988) Development 104, 7543 12 Hayashi, S. and Scott, M.P. (1990) Cell 63, 883-894 13 Gould, A.E, Brookman, J.L., Stmtt, D.I. and White, R.A.H. (1990) Nature 348, 308-312
REFERENCF~S 1 Robert.son, E.J. (1986) Trends Genet. 2, 9-13 2 Mansour, S.L., ~ m a s , K.R. and Capecchi, M.R. (1988) Nature 336, 348-352 3 Zimmer, A. and Gross, P. (1989) Nature338, 150-152 4 Robertson, E.J. (1987) in Teratocarcinomms and Embryonic Stem Cells- A Practical Approach (Robertson, E.J., ed.), pp. 71-112, IRL Press Contributed ~. S. E Chan and MJ. Evans, Department o f Genetics, University o f Camlm'dge. Cambn'dge CB2 3EI~. UK.
FIG MARCH1991 VOL. 7 XO. 3 -6
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developmental systems in other species, and enable the isolation of binding sites of other types of regulatory proteins. Acknowledgements l.J.J, is a Lister fellow. References 1 Garcia-Bellido, A. (1975) in Cell Patternitzg (Ciba Foundation Syrup. 29), pp. 161-178, Elsevier 2 Scott, MP., Tamkun, J.W. and Hartzell. G.W., III (1989) Biocbim Biophys. Acta 989, 25q8 3 Kissinger, C.R. etal. (1990) Cel163, ~75-590 4 lmmergluck, K., Lawrence, EA. and Bienz, M. (19~)) Cell62, 261-268 5 Reuter, R., Panganiban, G.E.F., Hoffman, F.M. and Scott, M.P.
(1990) Development 110, 1031-1040 6 Pollwein, 1:, Wagner, S. and Knippers, R. (1987) Nucleic Acids Res. 15, 9741-9759 7 Gruss, C. etal. (1988) Virolog)' 167, 349-360 8 Sompayrac, L. and Danna, K.J. (1990) Proc. Nat/A cad. Sci. I,X4 87, 3274-3278 9 Oliphant, A.R., Brandl, C0. and Struhl, K. (1989) Mol. Cell. Biol. 9, 2944-2949 10 Kinzler, K.W. and Vogelstein, B. (1990) Mol. Cell. Biol. 10, 634-642 11 Laughon, AP., Howell, W. and Scott, M.P. (1988) Development 104, 7543 12 Hayashi, S. and Scott, M.P. (1990) Cell 63, 883-894 13 Gould, A.E, Brookman, J.L., Stmtt, D.I. and White, R.A.H. (1990) Nature 348, 308-312
REFERENCF~S 1 Robert.son, E.J. (1986) Trends Genet. 2, 9-13 2 Mansour, S.L., ~ m a s , K.R. and Capecchi, M.R. (1988) Nature 336, 348-352 3 Zimmer, A. and Gross, P. (1989) Nature338, 150-152 4 Robertson, E.J. (1987) in Teratocarcinomms and Embryonic Stem Cells- A Practical Approach (Robertson, E.J., ed.), pp. 71-112, IRL Press Contributed ~. S. E Chan and MJ. Evans, Department o f Genetics, University o f Camlm'dge. Cambn'dge CB2 3EI~. UK.
FIG MARCH1991 VOL. 7 XO. 3 -6