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Single Step Guaranteed Knockdown No transfection reagents needed Ideal for cellular and animal studies
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AUM lnc

FANA-ASOs

FANA ASOs for
lncRNA Silencing and Regulation

Achieve potent RNase H-mediated cleavage of the target long non-coding RNA.

fna-fig-1

AUMlnc provides potent lncRNA 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), FANA single-stranded ASOs use RNase H-mediated cleavage (Figure 1). This mode of lncRNA knockdown is simpler than siRNA-mediated knockdown and eliminates RISC-associated off-target effects often observed with siRNA. Self-delivering FANA ASO can enter the nucleus and can be used to target both cytoplasmic and nuclear RNA.

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.

Case Studies

FANA-mediated lncRNA knockdown of EGFR-AS1 and MIR205HG
genes in Erlotinib sensitive cells (HCC 827 & NCI H2228)

RT-PCRT analysis showed a significant decrease in EGFR-AS1 (blue) and MIR205HG (red) expression in cells treated with FANA-EGFR-AS1 or FANA-MIR205HG compared to scrambled control. FANA-treated cells showed reduction in growth and higher proliferation rates. These in vitro experiments validated EGFR-AS1 and MIR205HG as determinants of Erlotinib response (Figure 2). Adapted from Nath, A. et al. 2019. PNAS.

Ordering information:

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

  • 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.
FANA vs shRNA

AUMlnc

FANA antisense oligonucleotides (FANA ASOs) for lncRNA silencing and regulation

AUMlnc provides potent lncRNA 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 lncRNA

  • High stability and resistance to endonucleases

FANA ASOs vs. siRNA
Not Required Transfection Reagents Required
High Efficiency in difficult-to-transfet cells Low-Moderate
Non-toxic Toxicity Can be toxic due to the use of transfection reagents
Easy and convenient one-step process Transition from cell culture to in vivo models Require extensie optimization use of delivery reagents
High: Resistant to nucleases Stability Moderate
No RISC-associated off-target effects Yes
FANAs have high binding affinity and specificity to the target RNA Specificity siRNA grade binding affinity and 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 Antisense Oligonucleotides (FANA ASOs) are uniquely modified with 2'-deoxy-2'-fluoro-arabinoguanosine (FANA) that enhances the intracellular stability of the oligos, as well as their binding to the target lncRNA. The FANA modifications also allow for the oligos to be self-delivered into cells without any transfection reagents, as well as in animals, without the need of special delivery formulations.

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.

Ordering information

Name Application Purification Study Model
AUMlnc lncRNA knockdown RPC Cell lines and primary cells
AUMlnc HPLC lncRNA knockdown Sensitive primary cells and animal models
AUMlnc In-vivo ready lncRNA knockdown Animal models
Notes:

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

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

To order please visit:
aumbiotech.com/RNAsilencing

Or email us at:

customercare@aumbiotech.com
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