In vitro Assays for Oligonucleotide Drug Discovery
In vitro screening of oligonucleotides is an essential tool in a drug discovery project. A properly executed in vitro screen can identify a shortlist of candidates to progress from a large oligonucleotide library, which can save on costly in vivo studies in the future. High throughput screening methods have greatly enhanced the workflow and timelines to help identify potent sequences efficiently with excellent reproducibility. In recent years, in vitro assays have also been developed to predict potential toxicity or immunogenicity and can be a quick supplement (or stand alone activity) for any screening cascade. These tox prediction studies can help flag any toxic or immunogenic oligonucleotides prior to in vivo testing, saving time and money.
In vitro Screening for Efficacy or Toxicity
Our dedicated team of oligonucleotide experts advise on the optimal parameters for your in vitro screening activities which are tailored to your needs. To deliver the highest quality data package to our partners, we take multiple parameters into consideration for our in vitro study designs, including cell line selection (iPSC-derived, primary or patient’s derived cells, 2D or 3D models), oligonucleotide chemistry, species, therapeutic area, desired outcomes, and timelines.
Examples of cells employed for in vitro profiling of oligonucleotides, ranging from iPSC-derived call models (either 2D or 3D) to patient-derived cells for splicing correcting oligonucleotides.
We can screen oligonucleotide libraries of up to 1,000 sequences in 384 well format by gymnosis or transfection with the appropriate assay controls for either initial screen, hit conformation CRC, or a long list of toxicity assays. The target engagement of your oligonucleotides will be validated at different levels (mRNA and protein) and, depending on your needs, on any disease-relevant phenotype associated with modulation of your target.
Our in vitro team can help with:
- Oligonucleotide library screening
- Immunogenicity assay with PBMC or BJAB cells
- Caspase 3/7 for hepatotoxicity prediction
- MEA assay for neurotoxicity predictions
- Multiple assay readouts including RT-qPCR, MCE (RNA) or Western blot/Simple Western (protein), RNAscope, RNAseq/transcriptomics, proteomics, metabolic, immune, FACS, IHC, IF, high content screening, cell painting, live imaging on IVIS
- Multiple cell models including 2D and 3D: cell lines, primary cells, patient’s derived cells, iPS cells, co-cultures, organoids, disease-relevant cells
- Mechanistic or delivery studies including receptor mediated delivery, testing conjugates, discovering ligands for targeted delivery and on-demand
