Bloom and Starr Labs SARS-CoV-2 RBD SSM Libraries
(Pooled Libraries #1000000182, #1000000183, #1000000184, #1000000185, #1000000186, #1000000187, #1000000188, #1000000231, #1000000232)
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Purpose
Site-saturation mutagenesis (SSM) libraries of yeast-displayed SARS-CoV-2 receptor-binding domain (RBD). Each SSM library is in the background of fixed mutations that define certain SARS-CoV-2 variants that have evolved, including the ancestral Wuhan-Hu-1 strain and variants Alpha (N501Y), Beta (K417N/E484K/N501Y), Delta (L452R/T478K), Eta (E484K), Omicron BA.1 (15 mutations), Omicron BA.2 (16 mutations), Omicron BQ.1.1 (20 mutations) and Omicron XBB.1.5 (23 mutations). Each library has a downstream N16 barcode that was previously linked to mutant identity via PacBio sequencing.
Libraries are useful for deep mutational scanning characterization of the impacts of SARS-CoV-2 mutations on receptor- and antibody-binding.
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Vector Backbone
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Depositing Labs
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Publication
#1000000182, #1000000183, #1000000184, #1000000185, #1000000186:
Starr et al Science. 2022 Jun 28:eabo7896. doi: 10.1126/science.abo7896.(How to cite )#1000000187, #1000000188:
Starr et al PLoS Pathog. 2022 Nov 18;18(11):e1010951. doi: 10.1371/journal.ppat.1010951.(How to cite )#1000000231, #1000000232:
Taylor et al. bioRxiv. 2023. doi: 10.1101/2023.09.11.557279. (How to cite )
Ordering
Item | Catalog # | Description | Quantity | Price (USD) | ||
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Pooled Library | 1000000182 | SARS-CoV-2 RBD Wuhan-Hu-1 SSM Library | 1 | $ 540 | Add to Cart | |
Pooled Library | 1000000183 | SARS-CoV-2 RBD Alpha SSM Library | 1 | $ 540 | Add to Cart | |
Pooled Library | 1000000184 | SARS-CoV-2 RBD Beta SSM Library | 1 | $ 540 | Add to Cart | |
Pooled Library | 1000000185 | SARS-CoV-2 RBD Delta SSM Library | 1 | $ 540 | Add to Cart | |
Pooled Library | 1000000186 | SARS-CoV-2 RBD Eta SSM Library | 1 | $ 540 | Add to Cart | |
Pooled Library | 1000000187 | SARS-CoV-2 RBD Omicron BA.1 SSM Library | 1 | $ 540 | Add to Cart | |
Pooled Library | 1000000188 | SARS-CoV-2 RBD Omicron BA.2 SSM Library | 1 | $ 540 | Add to Cart | |
Pooled Library | 1000000231 | SARS-CoV-2 RBD Omicron BQ.1.1 SSM Library | 1 | $ 540 | Add to Cart | |
Pooled Library | 1000000232 | SARS-CoV-2 RBD Omicron XBB.1.5 SSM Library | 1 | $ 540 | Add to Cart |
Library Details
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Mutagenized CDSSARS-CoV-2 Receptor Binding Domain
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Mutagenized region size603 bp
Each catalog item is a pair of duplicate libraries to be used separately as technical replicates, as described in (Starr and Greaney et al. 2020 Cell and Starr et al. 2022 bioRxiv).
Each catalog item also contains a separate bacterial stab of the parent control plasmid, as listed in the following table.
Pooled Library | Background | Parent Plasmid # | Plasmids/library |
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1000000182 | Wuhan-Hu-1 | 166782 | ~50,000 |
1000000183 | Alpha | 184405 | ~50,000 |
1000000184 | Beta | 184406 | ~50,000 |
1000000185 | Delta | 184407 | ~100,000 |
1000000186 | Eta | 184404 | ~50,000 |
1000000187 | Omicron BA.1 | 184408 | ~100,000 |
1000000188 | Omicron BA.2 | 184409 | ~100,000 |
1000000231 | Omicron BQ.1.1 | 208085 | ~100,000 |
1000000232 | Omicron XBB.1.5 | 208086 | ~100,000 |
Library Shipping
Each library will be delivered as two pooled DNA libraries in microcentrifuge tubes on blue ice (tubes labeled lib1 and lib2). The tube contents will not necessarily be frozen. For best results, minimize freeze-thaws.
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Volume∼10 µL
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Concentration25 ng/µL
You will also receive a bacterial stab of the parental-control plasmid.
Resource Information
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Protocols
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Depositor Data
- Table of primers (Bloom Lab Github) (Link opens in a new window)
- Table of codon variants [Wuhan-Hu-1, Alpha, Beta, Eta] (Bloom Lab Github) (Link opens in a new window)
- Table of codon variants [Delta] (Bloom Lab Github) (Link opens in a new window)
- Table of codon variants [Omicron BA.1, BA.2] (Bloom Lab Github) (Link opens in a new window)
- Table of codon variants [Omicron BQ.1.1, XBB.1.5] (Starr Lab Github) (Link opens in a new window)
- Representative Illumina sequencing results [Omicron BQ.1.1, XBB.1.5] (Starr Lab Github) (Link opens in a new window)
- Representative plasmid map [Omicron BQ.1.1] (GB, 31 KB)
- Representative plasmid map [Omicron XBB.1.5] (GB, 33 KB)
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Terms and Licenses
Depositor Comments
The following GitHub repositories are provided:
- SARS-CoV-2-RBD_DMS_variants (Link opens in a new window) Includes algorithms for analyzing the PacBio and Illumina sequencing data for the Wuhan-Hu-1, Alpha, Beta, and Eta libraries
- SARS-CoV-2-RBD_Delta (Link opens in a new window) Includes algorithms for analyzing the PacBio and Illumina sequencing data for the Delta library
- SARS-CoV-2-RBD_DMS_Omicron (Link opens in a new window) Includes algorithms for analyzing the PacBio and Illumina sequencing data for the Omicron BA.1 and BA.2 libraries
- SARS-CoV-2-RBD_DMS_Omicron-XBB-BQ (Link opens in a new window) Includes algorithms for analyzing the PacBio and Illumina sequencing data for the Omicron BQ.1.1 and XBB.1.5 libraries
The SARS-CoV-2 RBD Beta SSM Library is also described in Greaney et al. 2022 "A SARS-CoV-2 variant elicits an antibody response with a shifted immunodominance hierarchy" PLoS Pathog. doi: 10.1371/journal.ppat.1010248 (Link opens in a new window)
The SARS-CoV-2 RBD Delta SSM Library is also described in Greaney et al. 2022 "The SARS-CoV-2 Delta variant induces an antibody response largely focused on class 1 and 2 antibody epitopes" bioRxiv. doi: 10.1101/2022.03.12.484088 (Link opens in a new window)
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.
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For your Materials & Methods section:
For #1000000182, #1000000183, #1000000184, #1000000185, #1000000186, #1000000187, #1000000188:
SARS-CoV-2 RBD SSM Libraries were a gift from Jesse Bloom (Addgene # 100000018x)
For #1000000231, #1000000232:
SARS-CoV-2 RBD SSM Libraries were a gift from Tyler Starr (Addgene # 100000023x) -
For your References section:
For #1000000182, #1000000183, #1000000184, #1000000185, #1000000186, #1000000187, #1000000188
Shifting mutational constraints in the SARS-CoV-2 receptor-binding domain during viral evolution. Starr TN, Greaney AJ, Hannon WW, Loes AN, Hauser K, Dillen JR, Ferri E, Farrell AG, Dadonaite B, McCallum M, Matreyek KA, Corti D, Veesler D, Snell G, Bloom JD. Science. 2022 Jun 28:eabo7896. doi: 10.1126/science.abo7896. PMID 35762884.
For #1000000231, #1000000232:
Deep mutational scans of XBB.1.5 and BQ.1.1 reveal ongoing epistatic drift during SARS-CoV-2 evolution. Taylor AL, Starr TN. bioRxiv 2023. doi: 10.1101/2023.09.11.557279. bioRxiv 2023.09.11.557279 .