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Welcome!

This content was prepared for the 2022 C. elegans Conference on Development, Cell Biology, & Gene Expression. Madison, WI.

• Presenter: Erin Osborne Nishimura
• Date: August 12, 202
• Session: Single-Molecule Approaches, chaired by Benjamin Weaver

Please contact me if you have any questions or inquiries about the content!

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Slides

Here are the slides:

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Protocols

Many of the protocols I reference are documented in our latest methods paper:

Parker DM, Winkenbach LP, Parker A, Boyson S, Osborne Nishimura E (2021) Improved Methods for Single-Molecule Fluorescence In Situ Hybridization and Immunofluorescence in Caenorhabditis elegans Embryos. Curr Protoc. 2021 Nov;1(11):e299. doi: 10.1002/cpz1.299.

Because there is something wrong with the availability of that paper, I am including it here:

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References

Some of the manuscripts we cite in the talk are linked here. Click to download.

• Parker et al., 2020. mRNA localization is linked to translation regulation in the Caenorhabditis elegans germ lineage. Development. 147(13):dev186817. doi: 10.1242/dev.186817.
  • original manuscript on mRNA localization to P granules

• Parker et al., 2021. Improved Methods for Single-Molecule Fluorescence In Situ Hybridization and Immunofluorescence in Caenorhabditis elegans Embryos. Current Protocols. e299. doi: 10.1002/cpz1.299.
  • protocols covered in the workshopImmunofluorescence + smFISH
  • smiFISH
  • tips, tricks, & reagents
  • special checklist for troubleshooting

• Parker et al., 2022. It’s Just a Phase: Exploring the Relationship Between mRNA, Biomolecular Condensates, and Translational Control. Front Genet. 27;13:931220. doi: 10.3389/fgene.2022.931220.
  • review article on mRNA localization to various biomolecular condensates

• Winkenbach et al., 2022. Translation of the ERM-1 membrane-binding domain directs erm-1 mRNA localization to the plasma membrane in the C. elegans embryo. BioRXiV. doi: https://doi.org/10.1101/2022.05.11.491546
  • preprint on erm-1 mRNA to plasma membranes
  • describes performing smFISH after drug treatment

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2021 Presentation on single molecule mRNA live imaging

In 2021, I organized a Workshop on mRNA live imaging. I presented a talk to report where we were at the time on using MS2/MCP and PP7/PCP to image molecules of mRNA in living worms (embryos, gonads, & larvae). That talk is here:

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Join us!

A postdoctoral research position is available in the Osborne Nishimura Lab aimed at pursuing single-molecule resolution live imaging of mRNA transcripts! This is an exciting opportunity. Contact Erin to learn more.