Reproducible assembly of functional complexes at 10 mg siRNA scale
Assemblies less than 150 nm diameter
Report siRNA incorporation efficiency
Report on zeta potential and polydispersity siRNA incorporation efficiency
No aggregation in 50% mouse/human serum
Chemical stability data of the assembly for >30 days
Activity in Cell-based Assays
Greater than 50% reduction in target mRNA levels by target siRNA at concentrations <10 nM in media containing 10% serum
Less than 10% reduction in target mRNA levels by control siRNA at concentrations <10 nM in media containing 10% serum
>5-fold window between target gene silencing IC50 and IC50 for reduction in viability
Activity in at least 3 cell lines (i.e. HeLa, HepG2, Raw 264.7, or cell lines relevant to delivery system under evaluation)
If the delivery platform incorporates a targeting moiety evidence should be provided for targeting by 1) using cell lines with differential expression of targeted receptor and 2) using assemblies with "active" and "inactive or mutant" targeting moieties
In Vivo Performance in Mouse Models
Single i.v. dose at 1, 3 & 9 mg/kg for control and target siRNAs-containing delivery assemblies
If the delivery platform under evaluation incorporates a targeting moiety in vivo arms should evaluate the "targeted" as well as assemblies incorporating "inactive/mutant" targeting moieties
>50% reduction in target mRNA levels in target tissue at 1 mg/kg dose by target siRNA by 24-48 hr
<10% reduction in target mRNA levels in target tissue at 1 mg/kg dose by control siRNA by 24-48 hr
Demonstration of RNAi-mediated target mRNA cleavage by 5'-RACE
<10-fold cytokine induction (TNF?, IFN?, IL6) (2 and 24 hr) at 3 mg/kg
<10-fold increases in ALT and AST at at 3 mg/kg
No effect on body weight (48 hr)
Blood clinical chemistry and hematology data (48 hr)
Targeted Delivery
The nature of the targeting mechanism (passive or active) should be understood
In vitro, active targeting should be blocked with the appropriate soluble ligand or receptor
Receptor engagement should result in cellular internalization
Actively targeted materials should bind to the cell in the presence of endogenous levels of competing ligand (ex: transferrin, folate, galactosamine)
Controls for in vivo active targeting experiments should include a non-functional targeting moiety of similar class, molecular weight and pI (ex: irrelevant IgG)
Materials without a control targeting ligand are not good controls
Unrelated cell lines that do not express the receptor are not good controls
Gene silencing in target cells using actively targeted materials should be at least 5-fold greater than that observed with the negative targeting controls
To obtain our package of siRNA materials, protocols, and performance criteria please provide a non-confidential scientific overview of your opportunity via
Printer Friendly Page
