Matches in SemOpenAlex for { <https://semopenalex.org/work/W2950595002> ?p ?o ?g. }
- W2950595002 abstract "Abstract Background Intrabodies enable targeting of proteins in live cells, but it remains a huge task to generate specific intrabodies against the thousands of proteins in a proteome. We leverage the widespread availability of fluorescently labelled proteins to visualize and manipulate intracellular signalling pathways in live cells by using nanobodies targeting fluorescent protein tags. Results We generated a toolkit of plasmids encoding nanobodies against red and green fluorescent proteins (RFP and GFP variants), fused to functional modules. These include fluorescent sensors for visualization of Ca 2+ , H + and ATP/ADP dynamics; oligomerizing or heterodimerizing modules that allow recruitment or sequestration of proteins and identification of membrane contact sites between organelles; SNAP tags that allow labelling with fluorescent dyes and targeted chromophore-assisted light inactivation; and nanobodies targeted to lumenal sub-compartments of the secretory pathway. We also developed two methods for crosslinking tagged proteins: a dimeric nanobody, and RFP-targeting and GFP-targeting nanobodies fused to complementary hetero-dimerizing domains. We show various applications of the toolkit and demonstrate, for example, that IP 3 receptors deliver Ca 2+ to the outer membrane of only a subset of mitochondria, and that only one or two sites on a mitochondrion form membrane contacts with the plasma membrane. Conclusions This toolkit greatly expands the utility of intrabodies for studying cell signalling in live cells." @default.
- W2950595002 created "2019-06-27" @default.
- W2950595002 creator A5019399622 @default.
- W2950595002 creator A5055968941 @default.
- W2950595002 date "2019-02-08" @default.
- W2950595002 modified "2023-09-24" @default.
- W2950595002 title "A genetically-encoded toolkit of functionalized nanobodies against fluorescent proteins for visualizing and manipulating intracellular signalling" @default.
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