A synthetic defective interfering SARS-CoV-2
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Abstract
Viruses thrive by exploiting the cells they infect, but in order to replicate and infect other cells they must produce viral proteins. As a result, viruses are also susceptible to exploitation by defective versions of themselves that do not produce such proteins. A defective viral genome with deletions in protein-coding genes could still replicate in cells coinfected with full-length viruses. Such a defective genome could even replicate faster due to its shorter size, interfering with the replication of the virus. We have created a synthetic defective interfering version of SARS-CoV-2, the virus causing the Covid-19 pandemic, assembling parts of the viral genome that do not code for any functional protein but enable the genome to be replicated and packaged. This synthetic defective genome replicates three times faster than SARS-CoV-2 in coinfected cells, and interferes with it, reducing the viral load of infected cells by half in 24 hours. The synthetic genome is transmitted as efficiently as the full-length genome, suggesting the location of the putative packaging signal of SARS-CoV-2. A version of such a synthetic construct could be used as a self-promoting antiviral therapy: by enabling replication of the synthetic genome, the virus would promote its own demise.
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SciScore for 10.1101/2020.11.22.393587: (What is this?)
Please note, not all rigor criteria are appropriate for all manuscripts.
Table 1: Rigor
Institutional Review Board Statement not detected. Randomization not detected. Blinding not detected. Power Analysis not detected. Sex as a biological variable not detected. Cell Line Authentication not detected. Table 2: Resources
Experimental Models: Cell Lines Sentences Resources Primers and probes for the DI and DI0 genomes, for the SARS-CoV-2 genome, and for the ACTB gene of Vero-E6 cells, were labelled using the FAM dye, an IBFQ quencher and an additional internal (ZEN) quencher, and were synthetised by Integrated DNA Technologies. Vero-E6suggested: NoneSoftware and Algorithms Sentences Resources All work with the SARS-CoV-2 was conducted in Biosafety Level-3 conditions at the Eva J Pell Laboratory of Advanced … SciScore for 10.1101/2020.11.22.393587: (What is this?)
Please note, not all rigor criteria are appropriate for all manuscripts.
Table 1: Rigor
Institutional Review Board Statement not detected. Randomization not detected. Blinding not detected. Power Analysis not detected. Sex as a biological variable not detected. Cell Line Authentication not detected. Table 2: Resources
Experimental Models: Cell Lines Sentences Resources Primers and probes for the DI and DI0 genomes, for the SARS-CoV-2 genome, and for the ACTB gene of Vero-E6 cells, were labelled using the FAM dye, an IBFQ quencher and an additional internal (ZEN) quencher, and were synthetised by Integrated DNA Technologies. Vero-E6suggested: NoneSoftware and Algorithms Sentences Resources All work with the SARS-CoV-2 was conducted in Biosafety Level-3 conditions at the Eva J Pell Laboratory of Advanced Biological Research, The Pennsylvania State University, following the guidelines approved by the Institutional Biosafety Committee. Coinfection and RNA extraction: 200,000 transfected cells were seeded in each well of a 24-well plate (each well in triplicate), and incubated for 1 hour before being inoculated with SARS-CoV-2 at MOI=10. Advanced Biologicalsuggested: NoneA BLAST search revealed no off-target sequences neither in the SARS-CoV-2 nor in the Chlorocebus sabaeus genome. BLASTsuggested: (BLASTX, RRID:SCR_001653)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.
- No funding statement was detected.
- No protocol registration statement was detected.
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