Whenever generation of a complete gene knock-out (e.g. via CRISPR/Cas9) is either lethal or linked to induction of unfavourable compensatory mechanisms, a more regulated way of amending the function of a gene of interest (GOI) could be attained through RNA interference (RNAi). RNAi relies on engagement of endogenous RNA-induced silencing complex (RISC) and Argonaute (Ago) endonuclease to initiate sequence-specific degradation of a host’s mRNA, guided by small exogenously-introduced RNAs. We can generate knock-down of a GOI using either small interfering RNAs (siRNA) or small hairpin RNAs (shRNA). With siRNA we can quickly generate a transient reversible knock down of a GOI. Whereas shRNAs are suitable for lentiviral cloning, allowing stable and inducible integration into host’s genome, overcoming adverse effects of transient over-expression, while also lasting for longer.
o2h team can:
- Design siRNAs and shRNAs
- Perform transient or stable gene knock-down using lentiviral cloning
- Determine phenotype of knock-down cells with a choice of redouts (including gene-reporters, immuno-assays, imaging analysis)
- Validate targets and their engagement with small molecules
To know more about our biology services offering or to request our brochure, please reach out to us at discovery@o2h.com.
