In vitro oncology & cell line engineeringJack Crawford2020-02-18T13:27:56+00:00
In order to fully execute on the advantages of Hera’s SRG OncoRat, in vivo xenografts must be accompanied with optimized cell culture and gene editing. Genome edited cancer cell lines can be leveraged effectively to contribute to the drug discovery pipeline. For example, disease-linked mutations can be created or corrected, genes can be knocked out or edited for target identification.
Hera has developed a process specifically designed to improve cancer cell gene editing for downstream in vitro and in vivo oncology studies. One key component is the upfront optimization of cell protocols. Hera has the expertise to adopt or modify existing cell culture methods for any cell line. Click here to see a list of 75+ different cells we have worked with either in a cell engineering scope or in vivo xenograft studies.
VCaP cells were plated onto 96-well plates and allowed 72hours for adhering to plate. Cells were then adjusted to phenol-red-free RPMI 1640 with 5% CSS for another 48 hours. After 7days of incubation with Enz or Bic in agonist or antagonist mode (using synthetic androgen R1881), CellTiter-Glo Luminescent Cell Viability Assay (Promega) was performed using Cytation 3 Cell Imaging Multi-Mode reader (BioTek). Dose-response curves and percent viability of the samples in agonist and antagonist mode were calculated.
Results: Bic effectively induced proliferation of VCaP cells in dose-dependent manner, where as Enz decrease cell viability at higher doses. Both Bic and Enz antagonized the proliferative effect of synthetic androgen R1881.
Cancer cell gene editing case study
GSK knockout cell line project as published in The Journal of Biological Chemistry in 2015. Our client needed a cell model to explore regulators of innate immune response for their antiviral discovery program. We optimized the cell line and discovered 4 copies of the PDE12 gene target were present. Multiple gene editing tools we created and tested during our reagent optimization process. Several were selected to increase success of tetraploid knockout.
Outcome: successful gene editing in a difficult tetraploid HeLa target reveals that PDE12 degrades 2′,5′-oligoadenylate (2-5A), a secondary messenger involved in antiviral action of interferon.