Frequently Asked Questions

FANA ASO calculator and dilution recommendations

FANA technology is a 2'-deoxy-2'-fluoro-beta-D-arabinose sugar modification on single stranded antisense oligonucleotides that can be used for RNA silencing and regulation experiments.

FANA oligos can be self-delivered without the use of any delivery agent. FANAs have high intracellular stability as well as high binding affinity to the target RNA forming very stable heteroduplex structures. Unlike conventional technologies, FANAs have high fidelity for the RNA target and highly reduced off target effects (no RISC associated off target effects). These attributes represent a more potent, efficient and cost effective solution in the RNA silencing and regulation space. Next generation FANA RNA silencing and regulation technology can be used to very efficiently knockdown or regulate a variety of genetic modalities, like mRNA, microRNA and long non-coding RNA.

• mRNA knockdown (RNase H mediated knockdown)
• Regulation of microRNA. For example, as antagomirs for miRNA regulation
• Cleavage of microRNA
• Knock down of lncRNA and other non-coding RNA
• Exon skipping
• RNA splicing
• Translational control/blocking: Can act as steric blockers by hybridization with the target RNA and inhibit translation
• Can act as decoys by hybridization with the target RNA for genetic regulation
• Applications that require hybridization with nucleic acids especially RNA
• Can be used as probes for a variety of applications
• Can be used in clinical diagnostics for a variety of applications

FANAs have very high binding affinity and specificity for their RNA targets. FANAs can be used to degrade a wide variety of RNA targets (mRNA or lncRNA) using RNase H or act as steric blockers (miRNA inhibiton, translational control, exon skipping).

• AUMsilence – for mRNA knockdown
• AUMantagomir – for miRNA silencing
• AUMmirblocker – for miRNA regulation*
• AUMlnc – for long non-coding RNA knockdown
• AUMblock – for translational blocking
• AUMsplice – for RNA splicing experiments
• AUMskip – for exon skipping experiments
• AUMprobe – for probing experiments

*FANA oligos designed to bind with the miRNA binding site in the 3’UTR (acting as steric blocker) thus preventing miRNA binding to its target, eventually upregulating the target.

siRNAs are known to have off-target effects, need transfection reagents, and do not work well in animal or fish models. They are expensive and need optimization. FANA oligos can be self-delivered without the use of any delivery agent. Further, as opposed to double-stranded siRNAs that use the RNAi pathway (involving RISC), FANAs are single-stranded next generation antisense oligonucleotides that use RNase H to mediate cleavage of the target. This mode is much simpler and does not have RISC-associated off-target effects. Lastly, FANA oligos can go into the nucleus and can be used to knockdown nuclear targets as well.

shRNAs require lengthy cloning and optimization. They may also need to be delivered via viral vectors. This requires long preparation and optimization time , which can be expensive and time consuming. FANA oligos can be self-delivered without the use of any delivery agent. Further, as opposed to shRNAs that eventually use the RNAi pathway (involving RISC), FANAs are single-stranded next generation antisense oligonucleotides that use RNase H to mediate cleavage of the target. This mode is much simpler and does not have RISC associated off-target effects. Lastly, FANA oligos can go into the nucleus and can be used to knockdown nuclear targets as well.

CRISPR needs optimization and delivery reagents. CRISPR is well known to have a lot of off target effects (Unexpected mutations after CRISPR–Cas9 editing in vivo; Nature Methods 14, 547–548 2017). FANA ASOs can be self-delivered without the use of any delivery agent. Further, FANAs ASOs are made of next generation chemical modifications that enhances chemical and intracellular stability, as well as binding to the target RNA, forming very stable heteroduplex structures. Unlike conventional technologies, FANA technology also provides high fidelity for the RNA target and highly reduced off target effects (no RISC associated off target effects).

No. FANA oligos can be self-delivered without the use of any delivery agent.

FANA ASOs have a special chemical modification that allows them tobe easily taken up by the cells by gymnosis.

Yes. FANAs workwell in primary cells. FANA oligos can easily knockdown or regulate target RNA in hard to transfect cells (like primary cells, B-cells, T-cells, neurons etc.) and highly sensitive cells derived from patient samples.

Yes. FANAs can work in animal models.

Yes – FANAs can be used with transfection reagents.

Please review our protocol page to know the amount that may suit your need. Please visit www.aumbiotech.com/Protocols

For most cell types, RPC purification works. For very sensitive cells, HPLC purification is recommended. For animal studies, in vivo purification is recommended.

Yes. If you want FANA oligos to be fluorescently labeled at 5′ end or 3′ end kindly let us know your desired label(s) and we will provide you with a special quote.

We typically recommend ordering 3 FANA oligos per target (we guarantee knockdown for RNA silencing experiments). Optional – If you already have an oligo sequence that you want in FANA format, simply input it in an ACTG format.

We will design FANAs for your experiments. Just provide us the name of the species and Target ID (Gene ID, Official Symbol, Accession Number etc.).

We typically recommend ordering 3 FANA oligos per target (we guarantee knockdown for RNA silencing experiments).

If you have your own candidate sequences, we can most definitely convert them in FANA ASOs as well. We also recommend a combination of your sequences and our designed sequences.

Please visit www.aumbiotech.com/Protocols