PRESTO-Salsa Barcoded Reporter Plasmids
(Kit #
1000000292
)
Depositing Lab: Noah Palm
These 313 barcoded reporter plasmids cover nearly all conventional (non-olfactory) human G protein-coupled receptors (GPCRs), including both liganded and orphan receptors across multiple GPCR classes, enabling simultaneous screening of the near-complete GPCRome in a single well of a 96-well plate. When combined with the PRESTO-Tango GPCR kit (Addgene Kit #1000000068) it enables multiplexed GPCR bioactivity screening with limited sample volume.
This kit will be sent as bacterial glycerol stocks in 96-well plate format.
Original Publication
Highly multiplexed bioactivity screening reveals human and microbiota metabolome-GPCRome interactions. Chen H, Rosen CE, Gonzalez-Hernandez JA, Song D, Potempa J, Ring AM, Palm NW. Cell. 2023 Jul 6;186(14):3095-3110.e19. doi: 10.1016/j.cell.2023.05.024. Epub 2023 Jun 14. PubMed (Link opens in a new window) Article (Link opens in a new window)
Description
PRESTO-Salsa is a novel multiplexed GPCR screening technology (Chen et al., 2023 (Link opens in a new window)) that builds off of the Tango technology (Barnea et al., 2008 (Link opens in a new window)) and PRESTO-Tango technology (Kroeze et al., 2015 (Link opens in a new window)). Salsa leverages next-generation sequencing to enable simultaneous screening of all conventional GPCRs in a single well of a 96-well plate. In addition to increasing sample throughput, this technology allows evaluation of potential activation of all conventional GPCRs using precious small-volume samples (e.g., chemical fractionations from modest culture volumes or in vivo metabolomes).
Each PRESTO-salsa reporter plasmid is uniquely barcoded with a 10 bp DNA sequence. When co-transfected with a GPCR Tango plasmid into Salsa reporter cells (i.e., HEK 293T cells stably expressing a β-arrestin/TEV fusion protein), it can generate a GPCR reporter cell line in which GPCR activity can be quantified using the unique DNA barcode. By pooling 313 GPCR reporter cell lines and reseeding them into 96-well plates, one can perform multiplexed bioactivity screening with a single sample against 313 GPCRs simultaneously.
Figure 1: PRESTO-Salsa enables ultrahigh-throughput screening of nearly all conventional GPCRs. A) By pooling cells that clonally express specific GPCR-barcode pairs, PRESTO-Salsa enables simultaneous evaluation of 300+ GPCRs in a single tube via next generation sequencing (NGS). In this example, an in vitro cultured bacterial strain is producing a ligand for ‘GPCR2’ which leads to increased transcription of the corresponding GPCR2 barcode. B) Heatmap showing that PRESTO-Salsa accurately detects GPCR activation via barcode counting in pooled libraries. Activation of pooled reporter cells co-transfected with ~300 unique GPCR-barcode pairs were stimulated with known GPCR ligands or M. morganii bacterial supernatants containing GPCR ligands, and activation was determined via NGS and barcode counting. Select GPCRs are shown as proof-of-principle even though 313 conventional GPCRs were included in this experiment.
Kit Documentation
Protocols
PRESTO-Salsa Screening Protocol (DOCX, 25 KB)Additional supporting materials can be found in the original publication and in Song et al., 2026 (Link opens in a new window).
Web Applications
Palm Lab interactive web interface (Link opens in a new window): check whether your receptor of interest is covered and what it responded to.
Additional References
Commensal-derived acetylcholine enhances mucosal immune education. Song D, Duncan-Lowey B, Khetrapal V, Hamchand R, Deng T, Brown H, Wu A, Martin AL, Bauer KM, Zhao Y, Nguyen MT, Sonnert ND, Leopold SR, Wu Q, Crawford JM, Palm NW. Nature. 2026 Jul;655(8124):1027-1036. doi: 10.1038/s41586-026-10592-7. Epub 2026 Jun 3. PubMed (Link opens in a new window) Article (Link opens in a new window)
How to Cite this Kit
These plasmids were created by your colleagues. Please acknowledge the Principal Investigator, cite the article in which they were created, and include Addgene in the Materials and Methods of your future publications.
For your Materials and Methods section:
"The PRESTO-Salsa Barcoded Reporter Plasmids kit was a gift from Noah Palm (Addgene kit #1000000292)."
For your Reference section:
Highly multiplexed bioactivity screening reveals human and microbiota metabolome-GPCRome interactions. Chen H, Rosen CE, Gonzalez-Hernandez JA, Song D, Potempa J, Ring AM, Palm NW. Cell. 2023 Jul 6;186(14):3095-3110.e19. doi: 10.1016/j.cell.2023.05.024. Epub 2023 Jun 14. PubMed (Link opens in a new window) Article (Link opens in a new window)
PRESTO-Salsa Barcoded Reporter Plasmids - #1000000292
- Resistance Color Key
Each circle corresponds to a specific antibiotic resistance in the kit plate map wells.
- Inventory
Searchable and sortable table of all plasmids in kit. The Well column links to its kit plate map on the right, as well as lists the plasmid well location in its plate. The Plasmid column links to a plasmid's individual web page.
- Kit Plate Map
96-well plate map for plasmid layout. Hovering over a well reveals the plasmid name, while clicking on a well opens the plasmid page.
Resistance Color Key
| Ampicillin |