Identification of Pan‐Coronavirus Neutralizing Aptamers Through CRISmers Targeting the Conserved Fusion Peptide

Abstract A series of coronavirus outbreaks emphasize the necessity to develop pan‐coronavirus therapeutics. Previously, the CRISmers system is developed, a Clustered Regularly Interspaced Short Palindromic Repeats/CRISPR‐associated proteins (CRISPR/Cas)‐mediated cellular screening platform for the identification of RNA aptamers. In this study, CRISmers are applied to target the fusion peptide (FP) of Severe Acute Respiratory Syndrome Coronavirus 2 (SARS‐CoV‐2), a conserved region across coronavirus genera, with E.coli serving as the screening host. From this approach, a lead aptamer, #FP‐10 is identified, which demonstrates potent pan‐coronavirus neutralization activity against multiple SARS‐CoV‐2 variants and alphacoronaviruses (HCoV‐229E and HCoV‐NL63), demonstrating its potential for development into a broad‐spectrum therapeutic candidate.