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  Probing and manipulating intracellular membrane traffic by microinjection of artificial vesicles.

Koike, S., & Jahn, R. (2017). Probing and manipulating intracellular membrane traffic by microinjection of artificial vesicles. Proceedings of the National Academy of Sciences of the United States of America, 114(46), E9883-E9892. doi:10.1073/pnas.1713524114.

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Item Permalink: http://hdl.handle.net/11858/00-001M-0000-002E-1DE8-1 Version Permalink: http://hdl.handle.net/11858/00-001M-0000-002E-3511-3
Genre: Journal Article

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 Creators:
Koike, S.1, Author              
Jahn, R.1, Author              
Affiliations:
1Department of Neurobiology, MPI for biophysical chemistry, Max Planck Society, ou_578595              

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Free keywords: SNARE proteins; liposome; membrane traffic; microinjection; vesicle transport
 Abstract: There is still a large gap in our understanding between the functional complexity of cells and the reconstruction of partial cellular functions in vitro from purified or engineered parts. Here we have introduced artificial vesicles of defined composition into living cells to probe the capacity of the cellular cytoplasm in dealing with foreign material and to develop tools for the directed manipulation of cellular functions. Our data show that protein-free liposomes, after variable delay times, are captured by the Golgi apparatus that is reached either by random diffusion or, in the case of large unilamellar vesicles, by microtubule-dependent transport via a dynactin/dynein motor complex. However, insertion of early endosomal SNARE proteins suffices to convert liposomes into trafficking vesicles that dock and fuse with early endosomes, thus overriding the default pathway to the Golgi. Moreover, such liposomes can be directed to mitochondria expressing simple artificial affinity tags, which can also be employed to divert endogenous trafficking vesicles. In addition, fusion or subsequent acidification of liposomes can be monitored by incorporation of appropriate chemical sensors. This approach provides an opportunity for probing and manipulating cellular functions that cannot be addressed by conventional genetic approaches. We conclude that the cellular cytoplasm has a remarkable capacity for self-organization and that introduction of such macromolecular complexes may advance nanoengineering of eukaryotic cells.

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Language(s): eng - English
 Dates: 2017-10-30
 Publication Status: Published online
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 Table of Contents: -
 Rev. Method: Peer
 Identifiers: DOI: 10.1073/pnas.1713524114
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Title: Proceedings of the National Academy of Sciences of the United States of America
Source Genre: Journal
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Pages: - Volume / Issue: 114 (46) Sequence Number: - Start / End Page: E9883 - E9892 Identifier: -