Frequently asked Questions about Advirna's Self-Deliverable RNAi (sdRNA)

What is self-deliverable RNAi technology?

Self-deliverable RNAi technology provides a simple method of cell transfection for efficient and potent gene silencing while minimizing off-target effects. sdRNA technology is based on extensive chemical modification of siRNAs. This results in sdRNA’s stabilization and ability to penetrate cells without transfection reagents and/or harmful physical methods (e.g. electroporation). Learn more


Why use self-deliverable RNAi?

  • Transfections are easy with self-deliverable RNAi, simply dilute sdRNA into the cell media!
  • Self-deliverable RNAi can be used in all cell types, providing a solution for gene silencing in hard-to-transfect cells.

  • Artefacts associated with other transfection methods (i.e. lipid- mediated transfection) are eliminated with sdRNAi.

  • sdRNAs can be maintained in media for prolonged periods of time without any toxic effects or cell perturbations, allowing for passive transfection.

  • Knockdown of proteins with low turnover rates is more efficient and cost-effective. Unlike traditional knockdown with siRNAs, the stability of sdRNA makes it possible to maintain gene silencing without repeated transfections.

  • All of Advirna’s sdRNAs are validated for knockdown efficacy and efficacy in common cell lines, eliminating the guesswork of finding an effective solution for knockdown experiments.


How is Advirna’s sdRNA used?

Advirna's sdRNAs are provided as a ready-to-go solution that can be added directly to the culture media. See our product use guide for details.


Self-deliverable RNAi can be used with all cell types ex-vivo and in vivo. sdRNA is great for use in hard-to-transfect cell lines, such as primary neurons, hepatocytes, T-cells. Targeting proteins with long half-lives is easier with sdRNAs as they can be stably maintained in the cell media for long periods of time without toxic effects, and without need to add additional transfection reagents.

When can self-deliverable RNAi be used?


RNAi treatment results in knockdown of a specific gene expression on the mRNA level. With sdRNA reagents, RNAi knockdown typically results in over 70% knockdown as measured by qPCR. In some cases it is possible to achieve more than 90% knockdown on the mRNA level. The effect of knockdown is transient. The period of time depends on factors including cell division rate, siRNA stability, mRNA turnover, and target protein turnover rate. See our product guide for more info.

How effective is RNAi gene silencing?


Is sdRNA toxic to cells?

Our validated sdRNA is tested in-house for toxicity in HeLa cells at effective concentrations. At effective concentrations sdRNAs are not-toxic. Non-specific cellular toxicity can occur depending on the nature and conditions of the knockdown. Contact us with any questions regarding your specific application.


What is the longevity of knockdown induced by sdRNA?

In rapidly dividing cell lines (i.e. HeLa cells, T-cells) we observe knockdown effect (after removal of sdRNA from the media) with a 4-7 days duration, or approximately 8-10 cell divisions. In non-dividing cells (primary hepatocytes) sdRNA-induced knockdown can be observed for several weeks. Knockdown effect can be sustained by maintaining sdRNA in the cell media.


Are sdRNAs validated?

Yes, all of Advirna’s sdRNAs are validated using direct knockdown in standard cell lines (i.e. HeLa, PC12, etc.) or using a reporter system in HeLa cells.


Most potent sdRNAs are selected from a screening of several lead candidates selected using Advirna’s proprietory prediction algorithm. Lead candidates are screened for knockdown efficacy in established cell lines or using a reporter system if the gene of interest is only expressed in specialized cells (i.e. primary neurons). To decrease off-targeting, all sdRNAs are pre-screened for miRNA seed regions and designed against unique mRNA sequence sites (from RNAseq and RefSeq databases).

How are sdRNAs selected?

 

For more information on our product please see our Product Use Guide. Couldn’t find your answer here? Need further information or clarification? Get in touch and we can answer any additional questions.