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AUMsilence

FANA ASOs for mRNA Silencing

FANA ASOs for mRNA Silencing

Superior alternative to siRNA, shRNA & CRISPR. Self-delivery without transfection reagents.
High potency and low toxicity.

FANA ASOs for mRNA Silencing

AUMsilence provides potent mRNA knockdown by using FANA ASO technology, which is a third generation chemical modification platform. FANA ASO technology allows simple and efficient delivery into difficult-to-transfect cells including primary cells. Additionally, FANA ASOs can also be used for animal and preclinical studies without the need of transfection reagents or additional chemical formulations.

As opposed to the RNAi pathway (involving RISC machinery), FANA ASOs use RNase H-mediated cleavage (Figure 1). This mode of mRNA knockdown is simpler than siRNA-mediated knockdown and eliminates RISC-associated off-target effects often observed with siRNA. Unlike siRNAs that are processed in the cytoplasm, FANA ASO can enter the nucleus and can be used to target both cytoplasmic and nuclear RNA.

FANA antisense oligonucleotides (FANA ASOs) for mRNA knockdown and regulation

AUMsilence provides potent mRNA knockdown by using FANA technology which is a third generation chemical modification platform. FANA technology 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

FANA 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 FANA oligos with the cells Primary cells and difficult to transfect cells Easy to use. No transfection reagents required. Mix FANA oligos with the cells
Resistant to degradation by serum and cellular nucleases Stability Moderate stability
None RISC-associated off-target effects Yes
FANA proprietary 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.

FANA ASOs are uniquely designed and manufactured using 2′-deoxy-2′- fluoro-β-d-arabinonucleic acid (FANA) chemical modifications that enhance intracellular stability of the oligos, providing very high specificity as well as high binding affinity to the target mRNA. The FANA modifications also allow for the oligos to be self-delivered into cells without any transfection reagents or electroporation. FANA ASOs can be used for animal studies without the need of delivery formulations or conjugates.

Comparison of FANA ASO and siRNA mode of action

As opposed to the RNAi pathway (involving RISC) FANA 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, FANA oligos can go into the nucleus and can be used to target RNA present within the nucleus. Most importantly FANAs can be self delivered and do not need transfection reagents or delivery agents.

Case Studies

FANA-mediated knockdown of KCTD15 encoding gene in RS4;11 cells

FANA-treated cells compared with controls showed a progressive decrease in the KCTD15 mRNA level. KCTD15 is strongly upregulated in patients with B-cell acute lymphoid leukemia and in derived continuous cell lines (e.g., RS4;11)(Figure 2). Adapted from Smaldone, G. et al. 2019. Scientific Reports.

Knockdown of ABI1 by AUM’s ASOs in CD34+ isolated cells from bone marrow of healthy donors

ASOs penetrated 99% of cells and mediated depletion of ABI1 protein. A 2-fold increase in CD34+ cells were detected in S-phase, confirming the direct link between Abi-1 loss and cell cycle activity (Figure 3). Adapted from Chorzalska, A. et al. 2018. Blood.

Ordering information

Name Application Purification Study Model
AUMsilence mRNA knockdown RPC Cell lines and primary cells
AUMsilence mRNA knockdown HPLC Sensitive primary cells, animal models, and preclinical studies
AUMsilence mRNA knockdown In vivo Animal models and preclinical studies
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Notes:
  • Concentration: Please review our FANA ASO Calculation Guidelines.

  • Labeled FANAs: FANA ASOs can be labeled with any fluorescent label or tag (eg. biotin).

  • Duration of effect: Depending on the experiment, different time points can be used to measure inhibition or related effects for up to several days (and weeks in some cases) using a single dose or multiple doses. In certain cases (especially for very fast-growing cells), the knockdown effect may be reduced after a few days. In such cases, simply add more FANA ASOs to the cell culture to maintain knockdown levels.
  • Label: FANA ASOs can be labeled with any fluorescent label (red, green, yellow or others) to monitor cellular uptake or perform other biodistribution or probing experiments in animal studies. Additionally, FANA ASOs can also be labeled with any conventional tags (eg. biotin).
  • Storage: FANA ASOs are shipped in lyophilized form. Upon arrival, store them in -20°C. When ready to use, re-suspend FANA ASOs in sterile water or appropriate buffer at the desired concentration. Aliquot re-suspended FANA ASOs in aliquots to avoid multiple freeze-thaw cycles.
  • Size:FANA ASOs are available in 10, 25, 50, 100, 250, 500 and 1,000 nmoles. Higher amounts are also available upon inquiry.
To order please visit:
aumbiotech.com/RNAsilencing

Or email us at:

customercare@aumbiotech.com
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