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Cited by 603

The DOI matches its Crossref record; 5 of the 5 references it cites were found in a registry or an index.

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

Open access
Poser, Ina, Sarov, Mihail, Hutchins, James R A, Hériché, Jean-Karim, Toyoda, Yusuke, Pozniakovsky, Andrei, Weigl, Daniela, Nitzsche, Anja, Hegemann, Björn, Bird, Alexander W, Pelletier, Laurence, Kittler, Ralf, Hua, Sujun, Naumann, Ronald, Augsburg, Martina, Sykora, Martina M, Hofemeister, Helmut, Zhang, Youming, Nasmyth, Kim, White, Kevin P, Dietzel, Steffen, Mechtler, Karl, Durbin, Richard, Stewart, A Francis, Peters, Jan-Michael, Buchholz, Frank, Hyman, Anthony A·2008·Nature Methods

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.NOTE: In the version of this article initially published online, the name of one individual was misspelled in the Acknowledgments. The second sentence of the Acknowledgments paragraph should read, “We thank I. Cheesman for helpful discussions.” The error has been corrected for all versions of the article.

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Crossref · 10.1038/nmeth.1199

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Metadata

Authors
Poser, I., Sarov, M., Hutchins, J. R. A., Hériché, J.-K., Toyoda, Y., Pozniakovsky, A., Weigl, D., Nitzsche, A., Hegemann, B., Bird, A. W., Pelletier, L., Kittler, R., Hua, S., Naumann, R., Augsburg, M., Sykora, M. M., Hofemeister, H., Zhang, Y., Nasmyth, K., White, K. P., Dietzel, S., Mechtler, K., Durbin, R., Stewart, A. F., Peters, J.-M., Buchholz, F., & Hyman, A. A.
Venue
Nature Methods
Published
2008-04-06
Volume
5
Issue
5
Pages
409-415
ISSN
  • 1548-7091 (ISSN-L)
  • 1548-7105
OpenAlex
W1977416347
Cited by
603
References
5
FWCI
23.91
Type
journal article
Language
en
Open access
green

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