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New Technologies to Visualize Gene Expression with Single Molecule Resolution Live
Author(s) -
Stasevich Timothy
Publication year - 2022
Publication title -
the faseb journal
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.709
H-Index - 277
eISSN - 1530-6860
pISSN - 0892-6638
DOI - 10.1096/fasebj.2022.36.s1.0i197
Subject(s) - translation (biology) , messenger rna , computational biology , transcription (linguistics) , gene expression , gene , protein biosynthesis , biology , microbiology and biotechnology , genetics , linguistics , philosophy
My lab is creating technology to image and quantify gene activity in real time and with single‐molecule precision in living cells. Using new mRNA and protein tags, fluorescent probes, and single‐molecule microscopy, we image DNA transcription and mRNA translation live. In this talk, I will briefly introduce our technology and describe how it can be used to amplify fluorescence from newly synthesized proteins as they are being translated from single mRNAs. I will show how we quantify these signals to determine the size, shape, subcellular localization, and mobilities of mRNA translation sites, as well as their protein synthesis dynamics. I will then highlight some of our most recent advances, including (1) single‐molecule co‐imaging of transcription and translation of CRISPR‐edited endogenous genes, (2) optogenetic tools to tether mRNA during translation, (3) open‐source software to quantify single‐mRNA translation, and (4) a palette of genetically‐encoded fluorescent probes to light up translation in multiple colors with high fidelity.