Deciphering the RNA landscapes on mammalian cell surfaces

Area of Science:

• Cell Biology
• Molecular Biology
• Genomics

Background:

• Cell surface RNAs, including glycoRNAs, are known but their composition across cell types is poorly understood.
• Existing methods lack the specificity and sensitivity to comprehensively profile surface RNAs.

Purpose of the Study:

• To develop and validate a suite of methodologies for profiling, imaging, and quantifying cell surface RNAs.
• To characterize the diverse landscape of non-coding RNAs present on mammalian cell surfaces.

Main Methods:

• Developed AMOUR (Amplification-based Method for Unbiased RNA profiling) utilizing T7-based linear amplification.
• Integrated fluorescently labeled DNA probes with live cell imaging and flow cytometry.
• Utilized techniques to confirm membrane anchorage and quantify RNA abundance.

Main Results:

• Identified diverse non-coding RNAs on mammalian cell surfaces, extending beyond previously known glycoRNAs.
• Confirmed membrane localization and quantified abundance of specific RNAs including Y family RNAs, U5 snRNA, MTRNR2, MT-TA, VTRNA1-1, and XIST.
• Detailed the surface RNA landscapes of human umbilical cord blood mononuclear cells (hUCB-MNCs) and murine blood cells.

Conclusions:

• The developed methodologies provide effective approaches for investigating cell surface RNAs.
• The study expands the known repertoire of cell surface RNAs and provides a detailed characterization of their presence on specific cell types.
• This work lays the foundation for future research into the functional roles of cell surface RNAs.