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Cost-effective antisense, optimized for transfection

AUMsilence toASO

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A transfection-optimized antisense oligonucleotide, a gapmer with a pale central gap between its two wings, arrives in a lipid transfection particle: particles each carrying a gapmer drift outside the cell, one docks on the lip of a pit in the cell membrane and lets its strand through, and another strand leaves a vesicle inside. There one gapmer is paired with a messenger RNA with RNase H1, a translucent globular enzyme, on the gap under a warm glow; the messenger RNA is cut at the enzyme, with a clear gap in its strand before its beaded tail, and the oligonucleotide stays whole. One strand passes through a pore into the nucleus at the top right, and a mitochondrion lies below.

AUMsilence toASOs are antisense oligonucleotides (ASOs) for mRNA knockdown, delivered with a standard transfection reagent. In routine experiments in cell lines, and in screens across dozens or hundreds of targets, transfection is the delivery method already in use, and cost is a major consideration. A self-delivering oligonucleotide needs no transfection reagent and is used at a higher working concentration. A transfection-optimized antisense oligonucleotide is the same design as its self-delivering counterpart (AUMsilence sdASO, AUMantagomir sdASO or AUMlnc sdASO), with partial chemical modification, at a lower price. Complex it with the transfection reagent and add the complexes to the cells.

Transfection-optimized antisense oligonucleotides are built on the core strengths of the AUMsilence platform – specificity and stability – with chemical modifications calibrated for transfection protocols.

Sizes and purification

RPC
2 · 5 · 10 · 25 nmol
HPLC
2 · 5 · 10 · 25 nmol

Pools of four sequences against one target are available at 2, 5, 10 and 25 nmol per oligo (8, 20, 40 and 100 nmol in total).

Prices and lead time are shown on the order page. Custom sizes by quote.

At a glance

Class
Transfection-optimized antisense (toASO)
Targets
mRNA
Delivery
Deliver with a standard transfection reagent.
Fluorescent label
None on the target-specific oligo; labeled scramble controls in FAM, Cyanine 3, Cyanine 5 and Cyanine 7.

In the lab

The protocol in four steps

The steps below are the AUMsilence toASO protocol's, in its own words. The full protocol carries the amounts per plate format, the tips and the troubleshooting.

Read the full protocol
The protocol in cells, in four beats: plate the cells, prepare the stock, add the complexes to the cells, assess knockdownPlate cells: 50-70% confluency; Prepare the stock: 2 μM working stock; Add complexes to cells: 50-100 nM in the well; Assess knockdown: 24-72 hours.01Plate cells50-70% confluency02Prepare the stock2 μM working stock03Add complexes to cells50-100 nM in the well04Assess knockdown24-72 hours

  1. Step 1

    Cell preparation

    Plate cells in their optimum growth medium at a density appropriate for the cell type. A confluency of 50-70% at the time of transfection is typical.

  2. Step 2

    AUMsilence toASO stock preparation

    Prepare AUMsilence toASO stock solution by reconstituting lyophilized ASOs at the desired concentration. If you already have a stock solution prepared, skip to Step 3.

  3. Step 3

    Transfection method selection

    AUMsilence toASO can be delivered using various transfection methods. Choose the most appropriate method for your cell type and experimental needs.

  4. Step 4

    Analysis of knockdown efficiency

    Evaluate the knockdown efficiency of AUMsilence toASO at appropriate time points post-transfection.

Why

Features of AUMsilence toASO

  • Cost-effective for large experiments

    AUMsilence toASO is the AUMsilence sdASO design at a lower price, which makes it the choice for high-throughput studies or when budget is limited. A screen across dozens or hundreds of genes is the same approach at that price per oligonucleotide.

  • Flexible ordering options

    Choose between Individual toASOs for targeted gene silencing or a Pool of 4 toASOs. Our pool option provides 4 different toASO sequences targeting the same gene in one order.

  • Knockdown through RNase H1

    AUMsilence toASOs knock down the target RNA by RNase H1 cleavage, as AUMsilence sdASOs do. The combination of DNA-like and modified regions ensures they activate RNase H1 well and bind strongly.

  • High specificity, low off-target

    AUMsilence toASOs are sequence-specific, like our other products. They do not involve the RNA-induced silencing complex (RISC), so microRNA-like off-targeting does not occur. Any off-target would require a significant complementary match to the ASO itself, which our AI-driven design avoids by scanning the transcriptome.

  • Stable and nuclease-resistant

    AUMsilence toASOs are resistant to degradation by serum and cellular nucleases, because they keep the phosphorothioate backbone.

  • Multiple size options

    AUMsilence toASOs are available in 2, 5, 10, and 25 nmol, from a pilot study at 2 nmol to a large-scale experiment at 25 nmol, in RPC or HPLC purification.

Mechanism

Mechanism of action

RNase H1-mediated knockdown: AUMsilence toASOs are single-stranded DNA/RNA-mimicking oligonucleotides, similar in principle to our other products, that bind to target RNA and induce RNase H1 to cleave it. They are typically designed with partial chemical modification (a phosphorothioate backbone, not modified at every position), which reduces the synthesis cost. Once delivered into the cell (via a transfection reagent), they reach the nucleus and cytoplasm and seek out the target RNA. The mode of action is the same: formation of an ASO:RNA duplex, RNase H1 cleavage of RNA, and decreased target gene expression.

Requirement for delivery reagent: Unlike AUMsilence sdASOs, AUMsilence toASOs need a transfection reagent. Therefore, you will use standard transfection reagents or physical methods to introduce them. The AUMsilence toASO is mixed with a lipid nanoparticle or other carrier that facilitates its entry into cells. Once inside, however, no further differences exist – it acts as any AUM ASO does, by binding RNA.

Design and AI optimization: AUMsilence toASOs are still designed with our AI platform to provide optimal targeting of your gene of interest. They maintain a phosphorothioated backbone to resist nucleases and promote some gymnotic uptake, giving them an edge in stability over unmodified oligos. They are optimized to strike a balance: high activity, lower synthesis cost.

FigureHow AUMsilence toASO acts
Three stages inside a cell, left to right: a lipid transfection particle fused to the cell membrane releases a gapmer with a pale central gap between its two wings, while two more particles carrying gapmers drift in the medium outside; the gapmer pairs along its length with a messenger RNA between its capped start and its beaded tail, with no enzyme yet; RNase H1, a translucent globular enzyme, sits on the pale gap under a warm glow and the messenger RNA is cut beneath it into two clean ends, the gapmer whole across the cut, with short fragments of an earlier cut drifting below. The nucleus stands at the top right and a mitochondrion at the lower center.
  1. Stage 1

    The AUMsilence toASO is mixed with a lipid nanoparticle or other carrier that facilitates its entry into cells.

  2. Stage 2

    Once delivered into the cell (via a transfection reagent), AUMsilence toASOs reach the nucleus and cytoplasm and seek out the target RNA.

  3. Stage 3

    AUMsilence toASOs bind to target RNA and induce RNase H1 to cleave it.

Applications

Applications and use cases

  • High-throughput gene knockdown screens

    AUMsilence toASO is used for a screen across many genes (for example, a custom panel of 100 genes related to a pathway, to see which affects a phenotype). Plate cells in 96-well plates, transfect AUMsilence toASOs targeting each gene (plus controls), and read out effects.

  • Routine gene silencing in easy-to-transfect cells

    In cell lines like HeLa, HEK293, or other cancer cell lines that transfect with high efficiency, self-delivery is not needed. AUMsilence toASO is delivered to these cells with a standard transfection reagent.

  • Education and training

    AUMsilence toASO is used in academic teaching labs and in training students in gene silencing techniques, where budget is a concern. Students learn the antisense mechanism by transfecting cells with AUMsilence toASO and assessing gene knockdown.

  • Pilot experiments

    AUMsilence toASO is a first use of the AUMsilence platform at small scale: order it for the gene and test it with the transfection reagent already in use, in the assay already in use. The same target is then studied in primary cells or in vivo with AUMsilence sdASO, whose sequence can be the same.

Every compatible cell type, 3D model and organoid with the in vitro workflow, and in vivo with a delivery system.

See where it works

Experimental considerations

A transfection reagent is needed, and transfection reagents can introduce toxicity, alter the cellular response, or require additional optimization. In difficult-to-transfect cells, and in vivo without a carrier or formulation, AUMsilence sdASO is used instead.

The recommended working range for AUMsilence toASOs, delivered with a transfection reagent, is 50-100 nM. For AUMsilence sdASOs, which are self-delivering, the recommended range is 5-20 μM.

Ordering options

  • Individual toASO

    Perfect for targeted experiments

    • 2 nmol: Pilot studies
    • 5 nmol: Standard size
    • 10 nmol: Extended studies
    • 25 nmol: Scale-up

  • Pool of 4 toASO

    Four sequences against one target

    • 2 nmol × 4: 4 sequences, pilot
    • 5 nmol × 4: 4 sequences, standard
    • 10 nmol × 4: 4 sequences, extended
    • 25 nmol × 4: 4 sequences, scale-up

The toASO range

Three products and their controls, every one ordered from this page. Sizes are the delivered nmol. A pool of four is four sequences against one target at the stated amount per oligo.

The transfection-optimized products and their sizes
ProductRPCHPLCPool of four, RPC, per oligoLabeled, HPLC
AUMsilence toASO2, 5, 10, 25 nmol2, 5, 10, 25 nmol2, 5, 10, 25 nmol
AUMantagomir toASO2, 5, 10, 25 nmol2, 5, 10, 25 nmol2, 5, 10, 25 nmol
AUMlnc toASO2, 5, 10, 25 nmol2, 5, 10, 25 nmol2, 5, 10, 25 nmol
AUMscramble toASO2, 5, 10, 25 nmol2, 5, 10, 25 nmol10 nmol, FAM, Cyanine 3, Cyanine 5, Cyanine 7
AUMposctrl toASO2, 5, 10, 25 nmol2, 5, 10, 25 nmol

Product comparison

AUMsilence toASOAUMsiRNAAUMsilence sdASO
Delivery methodRequires transfectionRequires transfectionSelf-delivering
Best forBudget-conscious screeningRNA interference studiesPrimary cells and in vivo
Pool option✓ Pool of 4✓ PoolIndividual only
MechanismRNase H1RISC/Ago2RNase H1
In vivo studiesRequires carrier or formulationRequires carrier or formulationDirect injection

At the bench

Protocol and what arrives

Deliver with a standard transfection reagent. The protocol and the How much to order page are written for this product.

View the protocolHow much to orderHow to cite
Lead time
10-14 business days
Shipping
Products are shipped lyophilized, with priority shipping within the United States and international priority shipping elsewhere.
Quality control
Every oligo is strictly controlled for quality and is manufactured in an ISO 9001:2015 certified facility.

Related

Order AUMsilence toASO

Order your custom AUMsilence toASOs today. Simply provide us with your gene target information, and our AI-powered design system will create optimized oligos for your specific needs. We offer consultation on transfection protocols and experimental design for your gene silencing experiments.

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For research use only. Not for use in diagnostic or therapeutic procedures.