Whitehead SARS-CoV-2 Spike (S) Ectodomain and RBD Libraries
(Pooled Library #157971, #157972, #157973, #168776, #168777, #174293, #174294, #174295, #174296)
These libraries encode mutations to SARS-CoV-2 Spike ectodomain and receptor binding domain (RBD) that were screened in “Francino Urdaniz et al. One-shot identification of SARS-CoV-2 S RBD escape mutants using yeast screening”. The spike ectodomain and RBD sequences are codon optimized for S. cerevisiae.
Spike ectodomain mutations are encoded in the S1 (Library A, B) and S2 (Library C) regions. These libraries were used to screen for mutations that stabilize the ‘up’ prefusion conformation.
The RBD libraries include an N343Q mutation in order to make the RBD aglycosylated when expressed in S. cerevisiae. The RBD sequence has been codon-optimized for yeast expression, and encodes NNK mutations on surface residues. The depositing lab used these libraries to screen for escape mutants from a panel of monoclonal antibodies. The WT, N501Y, and E484Y libraries can be co-transformed with pJS697.
|Item||Catalog #||Description||Quantity||Price (USD)|
|Pooled Library||157971||SARS-CoV-2 Spike (S) Ectodomain library A||1||$350||Add to Cart|
|Pooled Library||157972||SARS-CoV-2 Spike (S) Ectodomain library B||1||$300||Add to Cart|
|Pooled Library||157973||SARS-CoV-2 Spike (S) Ectodomain library C||1||$300||Add to Cart|
|Pooled Library||168776||SARS-CoV-2 Spike (S) Receptor Binding Domain (RBD) library 1||1||$350||Add to Cart|
|Pooled Library||168777||SARS-CoV-2 Spike (S) Receptor Binding Domain (RBD) library 2||1||$350||Add to Cart|
|Pooled Library||174293||SARS-CoV-2 Spike (S) Receptor Binding Domain (RBD) library E484K Tile 1||1||$350||Add to Cart|
|Pooled Library||174294||SARS-CoV-2 Spike (S) Receptor Binding Domain (RBD) library E484K Tile 2||1||$350||Add to Cart|
|Pooled Library||174295||SARS-CoV-2 Spike (S) Receptor Binding Domain (RBD) library N501Y Tile 1||1||$350||Add to Cart|
|Pooled Library||174296||SARS-CoV-2 Spike (S) Receptor Binding Domain (RBD) library N501Y Tile 2||1||$350||Add to Cart|
|Library ID||Ectodomain Library||Spike region||Designed Mutations||Bacterial Resistance|
|Library ID||Receptor Binding Domain Library||Positions Covered||Designed Mutations||Bacterial Resistance|
|168776||WT RBD Library 1||333-436||1176||Kanamycin|
|168777||WT RBD Library 2||437-527||1323||Kanamycin|
|174293||E484K RBD Library 1||333-436||1120||Kanamycin|
|174294||E484K RBD Library 2||437-527||1260||Kanamycin|
|174295||N501Y RBD Library 1||333-436||1120||Kanamycin|
|174296||N501Y RBD Library 2||437-527||1260||Kanamycin|
This library is delivered as suspended DNA in a microcentrifuge tube on blue ice. The tube's contents will not necessarily be frozen. For best results, minimize freeze-thaws.
- Ectodomain Libraries (157971-157973): ∼10µL
- RBD Libraries (168776-168777,174293-174296): ∼20µL
- Ectodomain Libraries (157971-157973): 25ng/µL
- RBD Libraries (168776-168777,174293-174296): 50ng/µL
Terms and Licenses
The depositing lab recommends the following algorithm for analyzing NGS Data:: PACT (Github)
Improved mutant function prediction via PACT: Protein Analysis and Classifier Toolkit. Klesmith JR, Hackel BJ. Bioinformatics. 2019 Aug 15;35(16):2707-2712. doi: 10.1093/bioinformatics/bty1042. PubMed 30590444
These pooled libraries were created by your colleagues. Please acknowledge the Principal Investigator, cite the article in which the plasmids were described, and include Addgene in the Materials and Methods of your future publications.
For your Materials & Methods section:SARS-CoV-2 Spike library was a gift from Timothy Whitehead (Addgene #)
For your References section:One-shot identification of SARS-CoV-2 S RBD escape mutants using yeast screening. Francino-Urdaniz IM, Steiner PJ, Kirby MB, Zhao F, Haas CM, Barman S, Rhodes ER, Leonard AC, Peng L, Sprenger KG, Jardine JG, Whitehead TA. Cell Rep. 2021 Aug 31;36(9):109627. doi: 10.1016/j.celrep.2021.109627. Epub 2021 Aug 10. PubMed 34416153