HA Frankenbody Expression Plasmids

For Live Cell Imaging

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Inventors

Timothy Stasevich
Ning Zhao
Hiroshi Kimura
Yuko Sato

At A Glance

Researchers at Colorado State University have developed fluorescence microscopy tools to image proteins in living cells that are difficult if not impossible to see using standard techniques. They are using these techniques to answer long-standing questions about how epigenetic factors contribute to gene misregulation in human disease. Currently they are focusing on two hard-to-see protein populations: (1) nascent proteins in the process of being translated (while still attached to RNA) and (2) post-translationally modified proteins, particularly modified chromatin and modified transcription machinery.

These HA Frankenbody Expression Plasmids genetically encode scFvs that selectively bind the HA epitope (YPYDVPDYA) with high affinity in living cells and organisms and can be used to quantify HA-tagged protein translation, localization, and dynamics. The plasmids enable, state-of-the-art, single-molecule experiments in living cells, which we demonstrate by tracking single HA-tagged histones in U2OS cells and single mRNA translation dynamics in both U2OS cells and neurons.

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Licensing Director

Steve Foster
Steve.Foster@colostate.edu
970-491-7100

Reference No.:  18-093

HA Frankenbody Expression Plasmids
Plasmids encoding anti-HA single-chain variable fragments (scFv) fused with mEGFP, HaloTag, or mCherry under CMV promoter.

Highlights:

  • Encode mouse anti-HA- scFvs for H. sapiens (human) expression
  • Engineered scFv binding the linear HA epitope with high affinity and specificity in vivo
  • Light up in multiple colors HA-tagged nuclear, cytoplasmic, membrane, and mitochondrial proteins in diverse cell types
  • Precisely quantify the expression of HA-tagged proteins in live cells
  • Powerful tool for imaging protein dynamics in vivo
Publications

Zhao, N., Kamijo, K., Fox, P.D. et al. A genetically encoded probe for imaging nascent and mature HA-tagged proteins in vivo. Nat Commun 10, 2947 (2019). https://doi.org/10.1038/s41467-019-10846-1

Cecchetelli, Alyssa. “HA Frankenbody, a New Imaging Tool to Visualize Single Molecules and Nascent Peptides.” Addgene Blog Share Science, 10 Oct. 2019, https://blog.addgene.org/ha-frankenbody-a-new-imaging-tool-to-visualize-single-molecules-and-nascent-peptides.

Colorado State University. “It’s not an antibody, it’s a frankenbody: A new tool for live-cell imaging: Antibody-based probe works in living systems and targets the classic HA tag.” ScienceDaily. ScienceDaily, 3 July 2019. www.sciencedaily.com/releases/2019/07/190703121440.htm

Manning, Anne. “It’s Not an Antibody, It’s a Frankenbody: A New Tool for Live-Cell Imaging.” College of Natural Sciences, 3 July 2019, https://natsci.source.colostate.edu/its-not-an-antibody-its-a-frankenbody-a-new-tool-for-live-cell-imaging/.

Last updated: August 2021

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