sequence alignment c6 and rdrp (5 PRIME)

5 PRIME sequence alignment c6 and rdrp

Flowchart for the selection strategy. The selection criteria <t>and</t> <t>sequence</t> numbers remaining at the end of each stage were individually indicated. Vero E6 cells were transfected with 10 nM of siRNA before infection by SARS‐CoV‐2 at a multiplicity of infection (MOI) of 0.1 after 24 h. The numbers of viral RNA copies were quantitated with RT‐qPCR. The negative control siRNA served as the control and is abbreviated as “Ctrl.” C1–C11 represent the final candidate sequences after selection. The siRNAs capable of inhibiting up to 99% of viral envelope gene expression with P ‐values < 0.005 compared with Ctrl siRNA are marked with *. P ‐value by Student’s t ‐test. Vero E6 cells were transfected with 10 nM of siRNA before infection by SARS‐CoV‐2 at a MOI of 0.1 after 24 h. The number of infectious virions were quantitated with plaque‐forming assay. The siRNAs capable of inhibiting up to 99% of plaque‐forming virions production with P ‐values < 0.0001 compared with Ctrl siRNA are marked with *. P ‐value by Student’s t ‐test. IC 50 and sequences of modified siRNA C6G25S, C8G25S, and C10G31A. Vero E6 cells were transfected with 10, 2, 0.4, 0.08, or 0.016 nM of each modified siRNA and challenged with virus at MOI of 0.1. Plaque‐forming virions were detected 24 h after infection. The sequences and chemical modifications of sense (S) and antisense (AS) strand of each siRNA are presented with 2′‐F and 2′‐OMe modifications as green and black squares, respectively. No modified RNA residues and phosphorothioate interlinkages are depicted as white squares and red dots, respectively. IC 50 of C6 and the fully modified C6G25S. Vero E6 cells were transfected with 10, 2, 0.4, 0.08, or 0.016 nM of C6 or C6G25S before virus infection at an MOI of 0.1. The viral RNA was quantitated by RT‐qPCR at 24 h after infection. IC 50 data for viral <t>RdRp</t> inhibition by C6G25S. Vero E6 cells were transfected with 10, 2, 0.4, 0.08, or 0.016 nM of C6G25S before virus infection at an MOI of 0.1. The viral RNA was quantitated by RT‐qPCR at 24 h after infection. Data information: Data were analyzed with GraphPad Prism 5 software and presented as mean ± SD of three biological replicates in (B–F).

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Flowchart for the selection strategy. The selection criteria and sequence numbers remaining at the end of each stage were individually indicated. Vero E6 cells were transfected with 10 nM of siRNA before infection by SARS‐CoV‐2 at a multiplicity of infection (MOI) of 0.1 after 24 h. The numbers of viral RNA copies were quantitated with RT‐qPCR. The negative control siRNA served as the control and is abbreviated as “Ctrl.” C1–C11 represent the final candidate sequences after selection. The siRNAs capable of inhibiting up to 99% of viral envelope gene expression with P ‐values < 0.005 compared with Ctrl siRNA are marked with *. P ‐value by Student’s t ‐test. Vero E6 cells were transfected with 10 nM of siRNA before infection by SARS‐CoV‐2 at a MOI of 0.1 after 24 h. The number of infectious virions were quantitated with plaque‐forming assay. The siRNAs capable of inhibiting up to 99% of plaque‐forming virions production with P ‐values < 0.0001 compared with Ctrl siRNA are marked with *. P ‐value by Student’s t ‐test. IC 50 and sequences of modified siRNA C6G25S, C8G25S, and C10G31A. Vero E6 cells were transfected with 10, 2, 0.4, 0.08, or 0.016 nM of each modified siRNA and challenged with virus at MOI of 0.1. Plaque‐forming virions were detected 24 h after infection. The sequences and chemical modifications of sense (S) and antisense (AS) strand of each siRNA are presented with 2′‐F and 2′‐OMe modifications as green and black squares, respectively. No modified RNA residues and phosphorothioate interlinkages are depicted as white squares and red dots, respectively. IC 50 of C6 and the fully modified C6G25S. Vero E6 cells were transfected with 10, 2, 0.4, 0.08, or 0.016 nM of C6 or C6G25S before virus infection at an MOI of 0.1. The viral RNA was quantitated by RT‐qPCR at 24 h after infection. IC 50 data for viral RdRp inhibition by C6G25S. Vero E6 cells were transfected with 10, 2, 0.4, 0.08, or 0.016 nM of C6G25S before virus infection at an MOI of 0.1. The viral RNA was quantitated by RT‐qPCR at 24 h after infection. Data information: Data were analyzed with GraphPad Prism 5 software and presented as mean ± SD of three biological replicates in (B–F).

Journal: EMBO Molecular Medicine

Article Title: A siRNA targets and inhibits a broad range of SARS‐CoV‐2 infections including Delta variant

doi: 10.15252/emmm.202115298

Figure Lengend Snippet: Flowchart for the selection strategy. The selection criteria and sequence numbers remaining at the end of each stage were individually indicated. Vero E6 cells were transfected with 10 nM of siRNA before infection by SARS‐CoV‐2 at a multiplicity of infection (MOI) of 0.1 after 24 h. The numbers of viral RNA copies were quantitated with RT‐qPCR. The negative control siRNA served as the control and is abbreviated as “Ctrl.” C1–C11 represent the final candidate sequences after selection. The siRNAs capable of inhibiting up to 99% of viral envelope gene expression with P ‐values < 0.005 compared with Ctrl siRNA are marked with *. P ‐value by Student’s t ‐test. Vero E6 cells were transfected with 10 nM of siRNA before infection by SARS‐CoV‐2 at a MOI of 0.1 after 24 h. The number of infectious virions were quantitated with plaque‐forming assay. The siRNAs capable of inhibiting up to 99% of plaque‐forming virions production with P ‐values < 0.0001 compared with Ctrl siRNA are marked with *. P ‐value by Student’s t ‐test. IC 50 and sequences of modified siRNA C6G25S, C8G25S, and C10G31A. Vero E6 cells were transfected with 10, 2, 0.4, 0.08, or 0.016 nM of each modified siRNA and challenged with virus at MOI of 0.1. Plaque‐forming virions were detected 24 h after infection. The sequences and chemical modifications of sense (S) and antisense (AS) strand of each siRNA are presented with 2′‐F and 2′‐OMe modifications as green and black squares, respectively. No modified RNA residues and phosphorothioate interlinkages are depicted as white squares and red dots, respectively. IC 50 of C6 and the fully modified C6G25S. Vero E6 cells were transfected with 10, 2, 0.4, 0.08, or 0.016 nM of C6 or C6G25S before virus infection at an MOI of 0.1. The viral RNA was quantitated by RT‐qPCR at 24 h after infection. IC 50 data for viral RdRp inhibition by C6G25S. Vero E6 cells were transfected with 10, 2, 0.4, 0.08, or 0.016 nM of C6G25S before virus infection at an MOI of 0.1. The viral RNA was quantitated by RT‐qPCR at 24 h after infection. Data information: Data were analyzed with GraphPad Prism 5 software and presented as mean ± SD of three biological replicates in (B–F).

Article Snippet: The lower part of Fig shows the sequence alignment of C6 and RdRp , which is located in the 5‐prime region of ORF1b .

Techniques: Selection, Sequencing, Transfection, Infection, Quantitative RT-PCR, Negative Control, Control, Gene Expression, Modification, Virus, Inhibition, Software

Journal: EMBO Molecular Medicine

Article Title: A siRNA targets and inhibits a broad range of SARS‐CoV‐2 infections including Delta variant

doi: 10.15252/emmm.202115298

Figure Lengend Snippet: siRNA candidates against SARS‐CoV‐2.

Article Snippet: The lower part of Fig shows the sequence alignment of C6 and RdRp , which is located in the 5‐prime region of ORF1b .

Techniques: Sequencing

C6 targets a highly conserved region of the virus RdRp (accession number: NC_045512.2). The figure shows a genome map for five variants of concern (VOC), two variants of interests (VOI), and five other variants. Dots above the genome indicate the locations of typical mutations for each variant. Important amino acid mutations listed in the Spike mutations of interest on the European Centre for Disease Prevention and Control (ECDC) website are labeled in red. Other mutations are labeled in black. The target site and sequence for C6G25 recognition on RdRp is shown below the map. C6G25 sequence is shown in red and the viral sequence in black. IC 50 for C6G25S against different variants. Vero E6 cells were transfected with 10, 2, 0.4, 0.08, or 0.016 nM of C6G25S before infection with different strains of SARS‐CoV‐2. The viral RNA was quantitated by RT‐qPCR at 24 h after infection. Data are presented as mean ± SD of three biological replicates. Source data are available online for this figure.

Journal: EMBO Molecular Medicine

Article Title: A siRNA targets and inhibits a broad range of SARS‐CoV‐2 infections including Delta variant

doi: 10.15252/emmm.202115298

Figure Lengend Snippet: C6 targets a highly conserved region of the virus RdRp (accession number: NC_045512.2). The figure shows a genome map for five variants of concern (VOC), two variants of interests (VOI), and five other variants. Dots above the genome indicate the locations of typical mutations for each variant. Important amino acid mutations listed in the Spike mutations of interest on the European Centre for Disease Prevention and Control (ECDC) website are labeled in red. Other mutations are labeled in black. The target site and sequence for C6G25 recognition on RdRp is shown below the map. C6G25 sequence is shown in red and the viral sequence in black. IC 50 for C6G25S against different variants. Vero E6 cells were transfected with 10, 2, 0.4, 0.08, or 0.016 nM of C6G25S before infection with different strains of SARS‐CoV‐2. The viral RNA was quantitated by RT‐qPCR at 24 h after infection. Data are presented as mean ± SD of three biological replicates. Source data are available online for this figure.

Article Snippet: The lower part of Fig shows the sequence alignment of C6 and RdRp , which is located in the 5‐prime region of ORF1b .

Techniques: Virus, Variant Assay, Control, Labeling, Sequencing, Transfection, Infection, Quantitative RT-PCR

sequence alignment c6 and rdrp Search Results

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