BAC TransgeneOmics: a high-throughput method for exploration of protein function in mammals. Export

@article{citeulike:2694552, abstract = {The interpretation of genome sequences requires reliable and standardized methods to assess protein function at high throughput. Here we describe a fast and reliable pipeline to study protein function in mammalian cells based on protein tagging in bacterial artificial chromosomes (BACs). The large size of the BAC transgenes ensures the presence of most, if not all, regulatory elements and results in expression that closely matches that of the endogenous gene. We show that BAC transgenes can be rapidly and reliably generated using 96-well-format recombineering. After stable transfection of these transgenes into human tissue culture cells or mouse embryonic stem cells, the localization, protein-protein and/or protein-DNA interactions of the tagged protein are studied using generic, tag-based assays. The same high-throughput approach will be generally applicable to other model systems.}, address = {[1] Max Planck Institute for Molecular Cell Biology and Genetics, Pfotenhauerstrasse 108, D-01307 Dresden, Germany. [2] These authors contributed equally to this work.}, author = {Poser, I. and Sarov, M. and Hutchins, J. R. and H\'{e}rich\'{e}, J. K. and Toyoda, Y. and Pozniakovsky, A. and Weigl, D. and Nitzsche, A. and Hegemann, B. and Bird, A. W. and Pelletier, L. and Kittler, R. and Hua, S. and Naumann, R. and Augsburg, M. and Sykora, M. M. and Hofemeister, H. and Zhang, Y. and Nasmyth, K. and White, K. P. and Dietzel, S. and Mechtler, K. and Durbin, R. and Stewart, A. F. and Peters, J. M. and Buchholz, F. and Hyman, A. A.}, citeulike-article-id = {2694552}, citeulike-linkout-0 = {http://dx.doi.org/10.1038/nmeth.1199}, citeulike-linkout-1 = {http://dx.doi.org/10.1038/nmeth.1199}, citeulike-linkout-2 = {http://view.ncbi.nlm.nih.gov/pubmed/18391959}, citeulike-linkout-3 = {http://www.hubmed.org/display.cgi?uids=18391959}, day = {6}, doi = {10.1038/nmeth.1199}, issn = {1548-7105}, journal = {Nature methods}, month = {May}, number = {5}, pages = {409--415}, posted-at = {2009-02-23 22:48:07}, priority = {2}, title = {BAC TransgeneOmics: a high-throughput method for exploration of protein function in mammals.}, url = {http://dx.doi.org/10.1038/nmeth.1199}, volume = {5}, year = {2008} }

TY - JOUR ID - citeulike:2694552 L3 - citeulike-article-id:2694552 N2 - The interpretation of genome sequences requires reliable and standardized methods to assess protein function at high throughput. Here we describe a fast and reliable pipeline to study protein function in mammalian cells based on protein tagging in bacterial artificial chromosomes (BACs). The large size of the BAC transgenes ensures the presence of most, if not all, regulatory elements and results in expression that closely matches that of the endogenous gene. We show that BAC transgenes can be rapidly and reliably generated using 96-well-format recombineering. After stable transfection of these transgenes into human tissue culture cells or mouse embryonic stem cells, the localization, protein-protein and/or protein-DNA interactions of the tagged protein are studied using generic, tag-based assays. The same high-throughput approach will be generally applicable to other model systems. IS - 5 JF - Nature methods SN - 1548-7105 AD - [1] Max Planck Institute for Molecular Cell Biology and Genetics, Pfotenhauerstrasse 108, D-01307 Dresden, Germany. [2] These authors contributed equally to this work. EP - 415 TI - BAC TransgeneOmics: a high-throughput method for exploration of protein function in mammals. VL - 5 SP - 409 AU - Poser, I AU - Sarov, M AU - Hutchins, JR AU - Hériché, JK AU - Toyoda, Y AU - Pozniakovsky, A AU - Weigl, D AU - Nitzsche, A AU - Hegemann, B AU - Bird, AW AU - Pelletier, L AU - Kittler, R AU - Hua, S AU - Naumann, R AU - Augsburg, M AU - Sykora, MM AU - Hofemeister, H AU - Zhang, Y AU - Nasmyth, K AU - White, KP AU - Dietzel, S AU - Mechtler, K AU - Durbin, R AU - Stewart, AF AU - Peters, JM AU - Buchholz, F AU - Hyman, AA PY - 2008/05/06/ UR - http://dx.doi.org/10.1038/nmeth.1199 ER -