AI Article Synopsis
- Understanding the mRNA life cycle involves studying the dynamics and composition of mRNPs (messenger ribonucleoproteins), and techniques like fluorescence correlation spectroscopy (FCS) and cross-correlation spectroscopy (FCCS) are effective tools for this purpose.
- FCS can analyze mRNPs in cell lysates, which mimic live cell conditions and allow for detailed studies of mRNP interactions and responses to treatments.
- FCCS can confirm known RNA interactions and measure the overlap of molecules within mRNPs, enhancing our understanding of their complexity and providing new ways to quantitatively analyze these structures.
Article Abstract
Understanding the mRNA life cycle requires information about the dynamics and macromolecular composition and stoichiometry of mRNPs. Fluorescence correlation and cross-correlation spectroscopy (FCS and FCCS) are appealing technologies to study these macromolecular structures because they have single molecule sensitivity and readily provide information about their molecular composition and dynamics. Here, we demonstrate how FCS can be exploited to study cytoplasmic mRNPs with high accuracy and reproducibility in cell lysates. Cellular lysates not only recapitulate data from live cells but provide improved readings and allow investigation of single mRNP analysis under particular conditions or following enzymatic treatments. Moreover, FCCS employing minute amounts of cells closely corroborated previously reported RNA dependent interactions and provided estimates of the relative overlap between factors in the mRNPs, thus depicting their heterogeneity. The described lysate-based FCS and FCCS analysis may not only complement current biochemical approaches but also provide novel opportunities for the quantitative analysis of the molecular composition and dynamics of single mRNPs.
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| http://www.ncbi.nlm.nih.gov/pmc/articles/PMC8599746 | PMC |
| http://dx.doi.org/10.1093/nar/gkab751 | DOI Listing |
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