RNA Silencing in Insects

Oligos designed using next generation FANA technology can effectively and efficiently perform:
Efficient knock down or regulation of the target RNA.
Ability to bind to the target RNA (mRNA, miRNA or lncRNA) in a high sequence specific manner.
No toxicity.
Efficient delivery without an external source (e.g. without a transfection agent, formulation, conjugate or viral vector).

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  • Introduction

    mRNA Knockdown
    AUMsilence
    Key features: Superior alternative for other approaches. No delivery reagents required.
    AUMsilence self delivering oligos achieve highly efficient and potent antisense-based gene knockdown. No need to use toxic transfection reagents. Especially designed for primary cells and very difficult to transfect cell types. Ideal for animal studies. Suited for fish and amphibian model systems as well.
    Highly recommended as a replacement for siRNAs, shRNA, CRISPR or other oligo approaches.

    We take pride in having the simplest protocol for RNA silencing studies. FANA oligos can work with any cell type without causing toxicity. FANA can work with easy to transfect, hard to transfect (like primary cells, B-Cells, T-Cells, neurons etc.) and highly sensitive patient samples.

    Protocol:
    Just mix FANA oligos with the cells directly into the medium (without the use of transfection reagents).
    For RNA silencing (mRNA) experiments recommended working concentration is from 250 nM to 2.5 μM. It is recommended to perform a quick dose response using 2-3 working concentrations. Depending upon the cell type and RNA target, the knock down effect can be seen as early as 24 to 48 hours.

    miRNA Inhibition
    AUMantagomir
    Key features: High binding affinity. High specificity. No transfection reagents required. No toxicity
    AUMantagomir self transfecting oligos serve as potent antagomirs by binding with miRNAs and prevent their hybridization with their target mRNAs. AUMmirblocker self transfecting oligos serve as competitive blockers/decoys against miRNAs to inhibit their binding with target mRNA. AUMmirblocker oligos will bind with the targeted regions on the mRNA and inhibit binding of miRNA with the mRNA.
    We take pride in having the simplest protocol for RNA silencing studies. FANA oligos can work with any cell type without causing toxicity. FANA can work with easy to transfect, hard to transfect (like primary cells, B-Cells, T-Cells, neurons etc.) and highly sensitive patient samples.

    Protocol:
    Just mix FANA oligos with the cells directly into the medium (without the use of transfection reagents).
    For miRNA inhibition experiments recommended working concentrations are from 500 nM to 5 μM. It is recommended to perform a quick dose response using 2-3 working concentrations. Depending upon the cell type and miRNA target, the miRNA inhibition effect can be seen as early as 24 to 72 hours.

    Translation Blocking
    AUMblock
    Key features: No special conjugates or delivery vehicle required. No delivery reagents required.
    AUMblock oligos oligos can be used for to block translation experiments. This will not cause the degradation of the mRNA. We take pride in having the simplest protocol for such studies. No special conjugates or delivery vehicles required. This significantly reduces toxicity associated with delivery reagents.

    Highly recommended as a replacement for other conventional oligo approaches that cause toxicity.

    Description

    Depending upon the experiment different time points can be used to measure knock down or related effects for up to several days using a single dose (and weeks in some cases).
    In certain cases (especially for very fast growing cells) if the knock down effect is reduced after a few days, simply add more FANA oligos to the cell culture.
    FANA oligos can be fluorescently labeled (or with any desired label) to monitor cellular uptake or biodistribution. Still want to use transfection reagents: If, for any reason, you still prefer to use transfection reagents the recommended working concentration is from 1nM to 25 nM (depending upon the cell type).

    Shipping and Storage:
    FANA oligos are shipped in lyophilized form. Upon arrival store them in -20°C.
    When ready to use, mix FANA oligos with sterile water or your favorite buffer. It is recommended to make multiple aliquots to avoid multiple freez thaw cycles.

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