The AIRFoundry is equipped with state-of-the-art instruments to fulfill RNA-LNP orders and for users to access through our user programs. Our goal is to provide researchers with access to the latest technologies and expert support to accelerate their discoveries.
RNA Production
We perform in vitro transcription (IVT) reactions at a wide range of scales using a T7 RNA polymerase-based system. We fulfill large-scale requests for single RNAs (>10mg, up to 100mg) in addition to requests for small-scale productions of many RNAs (0.1-0.5mg each, such as for screening studies). Multiplex IVT is carried out efficiently by our Hamilton STAR robotic liquid handling system. Our RNA is column-purified to remove immunostimulatory double-stranded transcripts, and then subject to rigorous quality control analysis, including RNA concentration, purity (A260/A280 ratio), size and integrity (Agilent Bioanalyzer), and lack of immunostimlation (α-dsRNA dot blot and in vitro IFN release assay).
Unless otherwise requested, our RNA is N1-methylpseudouridine modified to ensure lack of innate immunogenicity. We also support a variety of other modifications, such as Cy3 or Cy5 fluorophores.
Prospective users typically initiate requests with an amino acid sequence which is codon-optimized by our team and cloned into a standard expression vector. Client-supplied plasmids may also be used for RNA production, with some restrictions (contact Facility Director for details).
Encapsulation of RNA into lipid nanoparticles (LNPs)
AIRFoundry formulates LNPs using two technologies. The Ignite NanoAssemblr™, allows for consistent and reproducible formulation of LNPs with tunable size and properties. LNPs formulated by our team are subject to characterization of size, polydispersity index, and encapsulation efficiency by our Stunner combinatorial analysis platform. Off-shelf LNPs expressing reporters such as GFP, mCherry, and luciferase are additionally tested for robust expression in vitro.
AIRFoundry uses the Infini HTFS system to automatically generate distinct RNA-LNP formulations for rapid screening of formulation conditions. RNA, aqueous buffers, lipids, lipid ratios, and N:P ratios can all be independently programmed, enabling up to 96 formulations to be generated directly from individual component stocks in a single run, with as little as 20 µg of RNA per sample. Formulations are automatically dispensed into well plates for further downstream analysis.
