AUM<i>silence</i> vs shRNA

AUMsilence Oligos

AUMsilence antisense oligos (AUMsilence ASOs) for mRNA knockdown and regulation

AUMsilence oligos use next-generation chemical modifications to achieve potent mRNA knockdown. AUMsilence oligos allows simple and efficient delivery into difficult-to-transfect cells and animals without the need of transfection reagents or formulations.

AUMsilence : Key features

  • Easy self-delivery without the need of transfection regents

  • Excellent uptake in difficult-totransfect cells and primary cultures

  • No toxicity

  • High specificity and affinity for target mRNA

  • High stability and resistance to endonucleases

AUMsilence ASOs vs. siRNA
Not Required Transfection Reagents Required
High Efficiency in difficult-to-transfect cells Low-Moderate
Non-toxic Toxicity Can be toxic due to the use of transfection reagents
Easy to use. No transfection reagents required. Mix AUMsilence oligos with the cells Primary cells and difficult to transfect cells Transfection reagents needed. May cause toxicity
Resistant to degradation by serum and cellular nucleases Stability Moderate stability
None RISC-associated off-target effects Yes
AUMsilence ASO technology allows very high specificity for the target RNA Specificity siRNA grade specificity

Lipid-based transfection and electroporation are widely utilized, conventional methods to deliver siRNA into the cells. However, in many primary cells, particularly immune cells, hematopoietic cells and neurons, lipid reagents and electroporation are associated with high toxicity and poor transfection efficiency. Alternative delivery methods, such as viral vectors, require laborious optimization and viral production steps, and carry associated risk of genome integration.

AUMsilence Antisense Oligonucleotides are modified with 3rd generation chemical modifications that enhances the intracellular stability of the oligos, as well as their binding to the target mRNA. The AUMsilence ASOs can be self-delivered into cells without any transfection reagents, as well as in animals, without the need of special delivery formulations.

Comparison of AUMsilence ASOs and siRNA mode of action

As opposed to the RNAi pathway (involving RISC) AUMsilence single-stranded antisense oligonucleotides use RNase H-mediated cleavage (Fig.1). This mode of mRNA knockdown is simpler than siRNA mediated knockdown and eliminates RISC-associated offtarget effects often observed with siRNA. Unlike siRNAs that are processed in the cytoplasm, AUMsilence oligos can go into the nucleus and can be used to target RNA present within the nucleus. Most importantly AUMsilence oligos can be self delivered and do not need transfection reagents or delivery agents.

Case Studies

Knockdown of Bcl-2 by ASOs in 518A2 melanoma cells and PC3 cells.

Densitometry analysis of western blot in 518A2 melanoma cells and PC3 cells after ASO treatment with varying concentrations showing up to 90% knockdown (Figure 2). Adapted from Souleimanian et al. 2012.

Knockdown of androgen receptor (AR) by ASOs in LNCaP prostate cancer cells, LAPC4 ad VCaP cells.

Densitometry analysis of western blot in in LNCaP prostate cancer cells, LAPC4 ad VCaP cells after oligo treatment with varying concentrations showing up to 90% knockdown (Figure 3). Adapted from Souleimanian et al. 2012.

Ordering information:

Product Name Purification Application Study model
AUMsilence RPC mRNA knockdown Cells lines and primary cells
AUMsilence HPLC mRNA knockdown Sensitive primary cells and animal models
AUMsilence In-vivo ready mRNA knockdown Animal models
Notes:
  • Labeled AUMsilence ASOsAUMsilence ASOs can be labeled with any fluorescent label or tag (eg. biotin).

  • Size: AUMsilence oligos are available in 10, 25, 50, 100, 250, 500 and 1000 nmoles. Higher amounts are also available.