Human MicroRNAs Interacting With SARS-CoV-2 RNA Sequences: Computational Analysis and Experimental Target Validation.

Area of Science:

• Virology
• Molecular Biology
• Genetics

Background:

• Severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) causes COVID-19, a global pandemic.
• MicroRNAs (miRNAs) are implicated in virus-host interactions, but their role in SARS-CoV-2 infection requires experimental validation.

Purpose of the Study:

• To identify and experimentally validate cellular miRNAs that target SARS-CoV-2 RNA sequences.
• To investigate the potential of these miRNA-viral interactions in understanding COVID-19 pathogenesis.

Main Methods:

• Bioinformatic prediction of cellular miRNAs targeting viral RNA.
• Experimental validation using luciferase reporter assays in A549 lung cells.
• Co-transfection with miRNA mimics to confirm interactions and assess impact on viral gene expression.

Main Results:

• Five of eight predicted cellular miRNAs showed interaction with viral sequences, reducing luciferase activity.
• Co-transfection with miRNA mimics further reduced luciferase activity, validating interactions with miR-219a-2-3p, miR-30c-5p, miR-378d, miR-29a-3p, and miR-15b-5p.
• miR-15b demonstrated repression of Spike gene expression; target sequences are conserved across SARS-CoV-2 variants.

Conclusions:

• Provides the first experimental evidence of cellular miRNA binding to SARS-CoV-2 sequences.
• Elucidates molecular mechanisms in SARS-CoV-2 infection and pathogenesis.
• Suggests potential for novel therapeutic and diagnostic tools targeting miRNA-viral interactions.