Intriguingly, an alternative but more dominant inhibitory mechanism of Remdesivir on SARS-CoV-2 RdRp has been proposed, in which Remdesivir shows stronger resistance to the NTP concentration increment when it was present at the template strand . In this scenario, the copy of RNA strand embedded with Remdesivir was used as the template and exerted the "template-dependent inhibition", where the UTP incorporation complementary to Remdesivir is inhibited. Interestingly, such inhibition can be significantly reduced by the point mutation V557L . These observations have altogether suggested that the presence of Remdesivir at the template strand offers the second opportunity to inhibit the RNA extension. However, the experimental assay alone is insufficient to distinguish whether the inhibition is exerted by directly disrupting the NTP incorporation at the active site or by hindering the translocation. Moreover, it remains elusive how V557L mutation renders the SARS-CoV-2 RdRp drug resistant to Remdesivir at the template strand.