Translation and Tethering (TnT) Plasmid

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. These techniques can visualize the translation of DNA, and answer long-standing questions about how epigenetic factors contribute to gene misregulation in human disease. Currently  these plasmids have been utilized on two hard-to-see protein populations such as: (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.

The “Translation and Tethering” TnT biosensor is a combination of the nascent chain tracking translation reporter and mRNA tethering reporters. The TnT biosensor available today is adaptable to any factor since tethering/recruitment is through a tethering cassette and not the endogenous RNA binding domain of a specific factor. TnT can be used to screen effects such as DNA translation as well as up and down regulation of proteins. Additionally, TnT can be used to study translation factors as well as effects of subcellular localization on translation by tethering TnT mRNA to a specific location.

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