BRET-Based Self-Cleaving Biosensors for SARS-CoV-2 3CLpro Inhibitor Discovery
This article has been Reviewed by the following groups
Listed in
- Evaluated articles (ScreenIT)
Abstract
The virus proteases 3CLpro are validated drug targets for developing antivirals to treat coronavirus diseases, such as COVID-19. However, the development of 3CLpro inhibitors relies heavily on BSL-3 laboratories.
Article activity feed
-
-
SciScore for 10.1101/2021.07.28.454072: (What is this?)
Please note, not all rigor criteria are appropriate for all manuscripts.
Table 1: Rigor
Ethics not detected. Sex as a biological variable not detected. Randomization not detected. Blinding not detected. Power Analysis not detected. Cell Line Authentication not detected. Table 2: Resources
Antibodies Sentences Resources The PVDF membrane was blocked with 1% (for anti-FLAG antibody) or 3% (for anti-HA antibody) non-fat milk in TBST buffer (Tris-buffered saline containing 0.05% Tween-20) overnight at 4 °C and incubated with anti-FLAG antibody (Sigma, F1804, 1:1000 dilution) or anti-HA antibody (Abcam, ab9110, 1:1000 dilution) for 2 hours at room temperature. anti-FLAGsuggested: (Sigma-Aldrich Cat# F1804, RRID:AB_262044)F1804suggested: (Sigma-Aldrich Cat# F1804, RRID:AB_262044)anti-HAsuggested: (Abcam …SciScore for 10.1101/2021.07.28.454072: (What is this?)
Please note, not all rigor criteria are appropriate for all manuscripts.
Table 1: Rigor
Ethics not detected. Sex as a biological variable not detected. Randomization not detected. Blinding not detected. Power Analysis not detected. Cell Line Authentication not detected. Table 2: Resources
Antibodies Sentences Resources The PVDF membrane was blocked with 1% (for anti-FLAG antibody) or 3% (for anti-HA antibody) non-fat milk in TBST buffer (Tris-buffered saline containing 0.05% Tween-20) overnight at 4 °C and incubated with anti-FLAG antibody (Sigma, F1804, 1:1000 dilution) or anti-HA antibody (Abcam, ab9110, 1:1000 dilution) for 2 hours at room temperature. anti-FLAGsuggested: (Sigma-Aldrich Cat# F1804, RRID:AB_262044)F1804suggested: (Sigma-Aldrich Cat# F1804, RRID:AB_262044)anti-HAsuggested: (Abcam Cat# ab9110, RRID:AB_307019)ab9110suggested: (Abcam Cat# ab9110, RRID:AB_307019)Experimental Models: Cell Lines Sentences Resources For western blot analysis, HEK 293T cells were lysed using RIPA buffer (Beyotime, P0013B). HEK 293Tsuggested: NoneRecombinant DNA Sentences Resources The DNA fragment encoding the biosensor pBRET-1 was synthesized at GENEWIZ (Suzhou, China) and inserted into pcDNA3.1 vector at the site after the FLAG-tag. pcDNA3.1suggested: RRID:Addgene_79663)The plasmid of pBRETmut-1 was constructed by introducing the 3CLpro C145A mutation into pBRET-1 through site-directed mutagenesis using primers C145A-F and C145A-R (Table S1). pBRETmut-1suggested: NonepBRET-1suggested: NoneSoftware and Algorithms Sentences Resources The DNA fragment encoding the biosensor pBRET-1 was synthesized at GENEWIZ (Suzhou, China) and inserted into pcDNA3.1 vector at the site after the FLAG-tag. GENEWIZsuggested: (GENEWIZ, RRID:SCR_003177)Results from OddPub: We did not detect open data. We also did not detect open code. Researchers are encouraged to share open data when possible (see Nature blog).
Results from LimitationRecognizer: An explicit section about the limitations of the techniques employed in this study was not found. We encourage authors to address study limitations.Results from TrialIdentifier: No clinical trial numbers were referenced.
Results from Barzooka: We did not find any issues relating to the usage of bar graphs.
Results from JetFighter: We did not find any issues relating to colormaps.
Results from rtransparent:- Thank you for including a conflict of interest statement. Authors are encouraged to include this statement when submitting to a journal.
- Thank you for including a funding statement. Authors are encouraged to include this statement when submitting to a journal.
- No protocol registration statement was detected.
Results from scite Reference Check: We found no unreliable references.
-