The DOI matches its Crossref record; 5 of the 5 references it cites were found in a registry or an index.
Rapid and efficient clathrin-mediated endocytosis revealed in genome-edited mammalian cells
Open accessAbstract
Clathrin-mediated endocytosis (CME) is the best-studied pathway by which cells selectively internalize molecules from the plasma membrane and surrounding environment. Previous live-cell imaging studies using ectopically overexpressed fluorescent fusions of endocytic proteins indicated that mammalian CME is a highly dynamic but inefficient and heterogeneous process. In contrast, studies of endocytosis in budding yeast using fluorescent protein fusions expressed at physiological levels from native genomic loci have revealed a process that is very regular and efficient. To analyse endocytic dynamics in mammalian cells in which endogenous protein stoichiometry is preserved, we targeted zinc finger nucleases (ZFNs) to the clathrin light chain A and dynamin-2 genomic loci and generated cell lines expressing fluorescent protein fusions from each locus. The genome-edited cells exhibited enhanced endocytic function, dynamics and efficiency when compared with previously studied cells, indicating that CME is highly sensitive to the levels of its protein components. Our study establishes that ZFN-mediated genome editing is a robust tool for expressing protein fusions at endogenous levels to faithfully report subcellular localization and dynamics.
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Citations
Cites
A Modular Design for the Clathrin- and Actin-Mediated Endocytosis Machinery
Kaksonen, M., Toret, C. P., & Drubin, D. G. (2005) · Cell · doi:10.1016/j.cell.2005.09.024 · Cited by 726 · cited 1×
“In yeast, direct genomic tagging of pairs of genes with different fluorescent tags has allowed researchers to define a very regular series of spatiotemporal events for CME in living cell”
Self-citationKuma, A., Matsui, M., & Mizushima, N. (2007) · Autophagy · doi:10.4161/auto.4012 · Cited by 555
Merrifield, C. J., Perrais, D., & Zenisek, D. (2005) · Cell · doi:10.1016/j.cell.2005.03.015 · Cited by 476
Enhancing zinc-finger-nuclease activity with improved obligate heterodimeric architectures
Doyon, Y., Vo, T. D., Mendel, M. C., Greenberg, S. G., Wang, J., Xia, D. F., Miller, J. C., Urnov, F. D., Gregory, P. D., & Holmes, M. C. (2011) · Nature Methods · doi:10.1038/nmeth.1539 · Cited by 402
Self-citationCargo- and adaptor-specific mechanisms regulate clathrin-mediated endocytosis
Mettlen, M., Loerke, D., Yarar, D., Danuser, G., & Schmid, S. L. (2010) · Journal of Cell Biology · doi:10.1083/jcb.200908078 · Cited by 177
Cited by
Molecular mechanism and physiological functions of clathrin-mediated endocytosis
McMahon, H. T., & Boucrot, E. (2011) · Nature Reviews Molecular Cell Biology · doi:10.1038/nrm3151 · Cited by 2,223
RNA-programmed genome editing in human cells
Jinek, M., East, A., Cheng, A., Lin, S., Ma, E., & Doudna, J. (2013) · eLife · doi:10.7554/elife.00471 · Cited by 2,098
Mechanisms of clathrin-mediated endocytosis
Kaksonen, M., & Roux, A. (2018) · Nature Reviews Molecular Cell Biology · doi:10.1038/nrm.2017.132 · Cited by 1,698
A guide to genome engineering with programmable nucleases
Kim, H., & Kim, J.-S. (2014) · Nature Reviews Genetics · doi:10.1038/nrg3686 · Cited by 1,163
Genome-editing Technologies for Gene and Cell Therapy
Maeder, M. L., & Gersbach, C. A. (2016) · Molecular Therapy · doi:10.1038/mt.2016.10 · Cited by 719
Metadata
- Authors
- Doyon, J. B., Zeitler, B., Cheng, J., Cheng, A. T., Cherone, J. M., Santiago, Y., Lee, A. H., Vo, T. D., Doyon, Y., Miller, J. C., Paschon, D. E., Zhang, L., Rebar, E. J., Gregory, P. D., Urnov, F. D., & Drubin, D. G.
- Venue
- Nature Cell Biology
- Published
- 2011-02-06
- Volume
- 13
- Issue
- 3
- Pages
- 331-337
- ISSN
- 1465-7392 (ISSN-L)
- 1476-4679
- OpenAlex
- W1980529461
- Cited by
- 231
- References
- 5
- FWCI
- 10.62
- Type
- journal article
- Language
- en
- Open access
- green
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