This library of plasmids codes for MS2 virus-like particles packaged with various S gene variants (Amino acid 319-869). This library is made available under the OpenMTA to enable the development and validation of low-cost diagnostic tools for rapidly identifying clinically relevant SARS-CoV-2 variants. Additional variants will be added to the library when necessary based on WHO/PHE surveillance data.
Upon successful QC of the final two variants the complete library will be available as a single kit from Addgene. Should this not be affordable for laboratories (especially in the Global South) then they can contact me directly and I will ship the materials free of charge.
Plasmids that make up this library:
- MS2_SC2_S_WT
- MS2_SC2_S_Alpha
- MS2_SC2_S_Beta
- MS2_SC2_S_Gamma
- MS2_SC2_S_Delta
- MS2_SC2_S_Zeta
- MS2_SC2_S_Eta
- MS2_SC2_S_Theta
- MS2_SC2_S_Kappa
- MS2_SC2_S_Lambda
- MS2_SC2_S_Mu [currently undergoing QC at Addgene]
- MS2_SC2_S_C.1.2 [currently undergoing QC at Addgene]
These plasmids have the following mutations that fall within amino acid 319 and 869 of the S protein:
| Plasmid | Mutations | Sequence |
|---|---|---|
| MS2_SC2_S_WT | - | genbank |
| MS2_SC2_S_Alpha | N501Y, A570D, P681H, T716I | genbank |
| MS2_SC2_S_Beta | E484K, N501Y, A701V | genbank |
| MS2_SC2_S_Gamma | K417T, E484K, N501Y | genbank |
| MS2_SC2_S_Delta | L452R, T478K, P681R | genbank |
| MS2_SC2_S_Zeta | E484K | genbank |
| MS2_SC2_S_Eta | E484K, Q677H | genbank |
| MS2_SC2_S_Theta | E484K, N501Y, P681H | genbank |
| MS2_SC2_S_Kappa | L452R, E484Q, P681R | genbank |
| MS2_SC2_S_Lambda | L452Q, F490S, T859N | genbank |
| MS2_SC2_S_Mu | R346K, K417N, E484K, N501Y, D614G, P681H | genbank |
| MS2_SC2_S_C.1.2 | Y449H, E484K, N501Y, D614G, H655Y, N679K, T716I, T859N | genbank |
For instructions on how to prepare the VLPs refer to our paper. I've noticed that NiCo21(DE3) cells work just as well as Rosetta 2(DE3)pLysS.
- Sanger Sequencing (github) (protocols.io)
- Additional workflows under development...
Jameel Fund