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Whitehead SARS-CoV-2 Spike (S) Ectodomain and RBD Libraries
(Pooled Library #157971, #157972, #157973, #168776, #168777, #174293, #174294, #174295, #174296)

  • Purpose

    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.

  • Vector Backbone
    Library ID Backbone
    SARS-CoV-2 Spike Ectodomain 157971-157973 pUC19
    SARS-CoV-2 Spike RBD (WT) 168776-168777 pJS699
    SARS-CoV-2 Spike RBD (N501Y) 174293-174294 pIFU001
    SARS-CoV-2 Spike RBD (E484K) 174295-174296 pIFU002

Ordering

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
Available to Academic and Nonprofits Only

Library Details

SARS-CoV-2 Spike Libraries
Library ID Ectodomain Library Spike region Designed Mutations Bacterial Resistance
157971 Ectodomain A SD1 1,468 Kanamycin
157972 Ectodomain B SD1 325 Ampicillin
157973 Ectodomain C SD2 138 Ampicillin
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

Library Shipping

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.

  • Volumes
    • Ectodomain Libraries (157971-157973): ∼10µL
    • RBD Libraries (168776-168777,174293-174296): ∼20µL
  • Concentrations
    • Ectodomain Libraries (157971-157973): 25ng/µL
    • RBD Libraries (168776-168777,174293-174296): 50ng/µL

Depositor Comments

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

bioRxiv 2021.03.15.435309.

How to cite this pooled library ( Back to top )

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