Genes with 5′ terminal oligopyrimidine tracts preferentially escape global suppression of translation by the SARS-CoV-2 Nsp1 protein

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FIGURE 6.
FIGURE 6.

mTOR inhibition affects Nsp1 mediated translation of TOP mRNA. (A) Matrix showing the experimental design for assessing the effect of Torin 1 or DMSO used as a vehicle control on EEF2 UTR bearing reporter constructs (TOP) or in constructs wherein the TOP motif of EEF2-UTR has been mutated (Mutant-TOP) in presence of Nsp1 or Nsp2. (B) The ratio of Renilla luciferase to Firefly luciferase activity for TOP and mutant-TOP reporters expressing HEK293T cells in presence of Nsp1 or Nsp2 and Torin 1 or DMSO. (C) Matrix showing the experimental design for assessing the effect of LARP1 on EEF2 UTR bearing reporter constructs (TOP) or in constructs wherein the TOP motif of EEF2-UTR has been mutated (Mutant-TOP) in presence of Nsp1 or Nsp2 in HEK293T-Cas9 (as control) or LARP1 knockout (KO) cell line. (D) The ratio of Renilla luciferase to Firefly luciferase activity for TOP and Mutant-TOP reporters expressed in HEK293T-Cas9 (as control) or LARP1 knockout (KO) cell line in presence of Nsp1 or Nsp2.

This Article

  1. Published in Advance June 14, 2021, doi: 10.1261/rna.078661.120 RNA 27: 1025-1045

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