Smart platform for siRNA screening & modification

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Smart platform for siRNA screening & modification

In recent years, many studies have focused on developing therapeutic agents using short interfering RNA (siRNA). The RNA interference technique using siRNA is a revolutionary method that can selectively suppress the expression of target genes.

EnhancedBio has established unique technologies to develop siRNA therapeutics including hit-lead-candidate finding and PK/TK/PD analyses. Moreover, we have developed a variety of tools to identify the level of RNA interference with siRNAs both in vitro and in vivo.

[References] • HS Jung and YK Shin. The potential RNAi-based combination therapeutics. Archives of pharmacal research. 34 (1), 1-2. doi:0.1007/s12272-011-
  0100-9 (2011)
• HS Jung et al. The synergistic therapeutic effect of cisplatin with Human papillomavirus E6/E7 short interfering RNA on cervical cancer cell lines in
  vitro and in vivo. International journal of cancer. 130 (8), 1925-1936. doi:10.1002/ijc.26197 (2011)

"Hit-to-Lead" candidate drug discovery is based on the process of establishing at least 10 siRNA sequence designs for the same target genes using siRNA library screening followed by selection of the highest quality lead siRNAs. in vivo efficacy of the selected lead siRNAs is then determined in experimental animal models.

in vitro / in vivo assay development
• siRNA physicochemical character ID (size, pH, N:P Ratio etc.)
• siRNA transfection protocol optimization
In vitro efficacy test (phenotype, cell death and aging, and cell cycle arrest)
• RT-qPCR (mRNA knock-down level)
• Western blotting (protein knock-down level)
• Animal experiment design (lead validation)
Chemical modification
Various modifications can be made to enhance in vivo stability.
Stable derivatives
Sense (S) and antisense (AS) strand are partially modified and combined.
Lead optimization
  • • Verification of the safety of siRNA derivatives in human serum is performed using the serum stability test.
  • • Analysis of siRNA derivatives superior to the lead substances using advanced assays, RT- qPCR, western blotting is conducted for
       validation of mRNA/protein level changes.
  • • Optimized siRNA candidate selection and animal experiments using candidate substances are performed.
Chemical modification
The siRNA pool is designed to reduce the off-target effect by using a lower concentration than the previously used concentration and to demonstrate superior therapeutic efficacy.