We apply advanced genetic engineering approaches to precisely modify the host genome in order to investigate the function of a gene of interest (GOI). In a basic application, selected guide sequences can direct a nuclease to a specific genomic site within a GOI, where targeted modifications introduced via nonhomologous end joining (NHEJ) result in gene disruption. If the knock-out is viable, its phenotype can be compared with that of the wild-type counterpart. This comparison provides insight into the consequences of loss of function and helps validate a target’s involvement in the studied cellular pathway or disease model. Gene knock-out can also confirm engagement of a small molecule with its target and identify potential off-target effects.
Furthermore, combining a guide sequence with a DNA repair template and leveraging homology-directed repair (HDR) enables precise modification of defined amino acid residues with putative functional roles. This allows refinement of the final gene product, system phenotype, or specificity of small-molecule binding.
We can introduce indels, point mutations, tags, and protein markers through genome engineering using NHEJ or HDR mechanisms to validate target function and small-molecule engagement in cells, supported by a range of downstream readouts (including reporter assays, immunoassays, and imaging analysis).
o2h team can:
- Design guide sequences and DNA repair templates
- Perform genome editing in mammalian cells and generate clonal populations
- Assess the phenotype of knock-out or knock-in cells using a range of readouts (reporter assays, immunoassays, imaging analysis)
- Validate targets and their engagement with small molecules
- Conduct functional mutagenesis analysis of endogenous amino acids
- Introduce tags and markers onto endogenous proteins for downstream biochemical, biophysical, or imaging applications
To know more about our biology services offering or to request our brochure, please reach out to us at discovery@o2h.com.
