The only self-delivering siRNA for difficult-to-transfect cells carries a functional guarantee!
Accell siRNA provides a revolutionary solution for RNAi in cells that are resistant to conventional transfection methods, or cannot tolerate viral transduction or electroporation. Hundreds of peer-reviewed publications have demonstrated successful target gene silencing in primary neurons, β-islet cells, macrophages, T-cells, and more, including in vivo studies.
The main determiner in successful delivery (and subsequent silencing) by Accell siRNA is the cells’ tolerance to serum-free conditions, since serum interferes with Accell siRNA’s passive delivery. If the following three steps are carried out, we guarantee at least 75% silencing (measured by qPCR) by the Accell SMARTpool and three of the Set of 4 individual siRNAs targeting your gene of interest.
Serum-free medium can be:
Evaluate the cells’ overall appearance and viability in comparison to cells grown for 72 hours in their normal growth medium. If the serum-free cells do not have significant differences in observable characteristics (viability, growth rate, morphology) to those in growth medium, proceed to Step 2. If not, contact technical support for assistance.
Culture cells in serum-free medium (as in Step 1) with positive and negative (non-targeting) controls at 1 µM final siRNA concentration, along with untreated cells. Recommended controls (Table 1) are available individually or as Accell Control Kits (red: Cat # K-005000-R1-01, -02, -03 or green: Cat # K-005000-G1-01, -02, -03).
The full Accell Delivery Protocol can be found here.
Table 1: Accell Positive and Negative Controls
At 72 hours, carry out the following assessments:
Perform all the same comparisons as in Step 2, but include mRNA level quantitation for your gene(s) of interest. When compared to Non-targeting siRNA, you should see at least a 75% reduction. If you do not, contact technical support.
If all three of the steps above are carried out as described, you are guaranteed at least 75% target gene silencing by either the SMARTpool or three of the Set of 4 individual siRNAs. If, after contacting technical support and carrying out recommended troubleshooting, your target gene is not silenced by at least 75%, a one-time replacement of the affected Accell SMARTpool or Set of 4 siRNA reagents will be provided.
Author: Louise C. Baskin, Senior Product Manager at Dharmacon
List of peer-reviewed citations of successful Accell siRNA application
Delivery of Accell siRNA to adherent or suspension cells
Repeated dosing resulted in knockdown up to 30 days
An overview of good experimental planning and design using Accell siRNA