miRNA inhibition oligos

AUMantagomir sdASO

Name: AUMantagomir sdASO
Brand: AUM BioTech

Self-delivering antisense oligonucleotides for microRNA inhibition

A self-delivering antisense oligonucleotide, one uniform strand with no gap, is taken up by endocytosis and binds a microRNA: single strands drift in the medium outside the cell membrane with no transfection reagent, the membrane takes one in through a pit and a vesicle, one passes through a pore into the nucleus, and in the cytoplasm one is paired end to end with a short microRNA, holding it in a duplex under a warm glow. Nothing is cut: the oligonucleotide and the microRNA both stay whole, no enzyme sits on the pair, and to the right of it an Argonaute protein, a translucent two-lobed body with an open groove, sits empty above a mitochondrion.

AUMantagomir sdASOs are self-delivering antisense oligonucleotides (ASOs) for microRNA (miRNA) inhibition in cell lines and difficult-to-transfect primary cells. MicroRNAs are small regulatory RNAs that typically bind to mRNAs and repress their translation or cause degradation. AUMantagomir sdASOs act as "antagomirs" – they bind to a specific microRNA of interest and prevent it from interacting with its natural mRNA targets.

Many microRNA studies are done in primary neurons, cardiomyocytes and tissue explants, which are difficult to transfect with conventional methods. Transfection reagents, electroporation, or viral vectors can introduce toxicity, alter the cellular response, or require additional optimization. Built on the same AUMsilence platform as our other products, AUMantagomir sdASOs enter the cell without a transfection reagent, electroporation, or a viral vector. They are taken up by endocytosis and reach the nucleus as well as the cytoplasm. Once inside the cell, an AUMantagomir sdASO binds its complementary microRNA, blocking its function and often leading to its degradation.

MicroRNA inhibition and target de-repression
AUMantagomir sdASOs inhibit the target microRNA, and the genes it represses are de-repressed.
No transfection required
AUMantagomir sdASOs are added directly to cell culture, with no transfection reagent, electroporation, or viral vector.
Minimal toxicity
AUMantagomir sdASO shares the low toxicity profile of our other ASOs.
High binding affinity to microRNA
The chemical modifications in AUMantagomir sdASO confer high binding affinity to RNA targets.

Sizes and purification

RPC: 10 · 25 · 50 · 100 nmol
HPLC: 25 · 50 · 100 nmol
Labeled, HPLC: 25 nmol · FAM, Cyanine 3, Cyanine 5, Cyanine 7

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

At a glance

Class: Self-delivering antisense (sdASO)
Targets: miRNA inhibition
Delivery: Add to culture medium or inject in vivo.
Fluorescent label: FAM, Cyanine 3, Cyanine 5 or Cyanine 7

In the lab

The protocol in four steps

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

Read the full protocol

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 treatment is typical.
Step 2
AUMantagomir sdASO stock preparation
Prepare AUMantagomir sdASO stock solution by reconstituting lyophilized ASOs at the desired concentration. If you already have a stock solution prepared, skip to Step 3.
Step 3
AUMantagomir sdASO delivery to cells
Add AUMantagomir sdASO to the cells at the desired final concentration. The recommended working range is 5-20 μM, with a starting concentration of 10 μM. The optimal concentration varies with the target gene, the RNA class (messenger RNA, microRNA or long non-coding RNA) and the cell type, and should be determined by titration for each system.
Step 4
Incubation and analysis
Incubate cells with AUMantagomir sdASO and analyze miRNA inhibition at appropriate time points.

Step 1, prepare cells and the oligonucleotide: a clear 24-well plate of pink medium, a microtube of pale blue solution with its lid open and a pipette tip reaching into its mouth, and three short beaded violet strands drawn in the air above the tube, the slide's motif for the oligonucleotide. — For research use only. Not for use in diagnostic or therapeutic procedures.
Sources
The pictures are the object pictures of AUM BioTech's workflow slide, rendered views of the objects on a white ground; none is a photograph of an experiment and none records a measurement.
Tile 04 is a schematic of the three readouts, not data: the two conditions are drawn with a modest difference to show what each readout is, and no magnitude is asserted.

Step 2, add to the culture medium: a single-channel micropipette with a violet plunger held over a 24-well plate of pink medium, its tip in one well. — For research use only. Not for use in diagnostic or therapeutic procedures.
Sources
The pictures are the object pictures of AUM BioTech's workflow slide, rendered views of the objects on a white ground; none is a photograph of an experiment and none records a measurement.
Tile 04 is a schematic of the three readouts, not data: the two conditions are drawn with a modest difference to show what each readout is, and no magnitude is asserted.

Why

Features of AUMantagomir sdASO

MicroRNA inhibition and target de-repression
AUMantagomir sdASOs inhibit the target microRNA, and the genes it represses are de-repressed. In rat hippocampal slices, an AUMantagomir sdASO against miR-134-5p lowered the microRNA, and in slices exposed to amyloid beta it restored CREB-1 and BDNF expression.
No transfection required
AUMantagomir sdASOs are added directly to cell culture, with no transfection reagent, electroporation, or viral vector. Primary tissue explants and organotypic slices are treated the same way.
Minimal toxicity
AUMantagomir sdASO shares the low toxicity profile of our other ASOs. Traditional miRNA inhibitors often require high concentrations or complex formulations that can cause cell stress. By avoiding transfection reagents and using a chemistry optimized for biocompatibility, AUMantagomir sdASO ensures that any observed effects are due to miRNA inhibition, not off-target toxicity.
High binding affinity to microRNA
The chemical modifications in AUMantagomir sdASO confer high binding affinity to RNA targets. This means even a low-abundance microRNA can be captured by an AUMantagomir sdASO. How much of the microRNA is held away from its natural targets depends on the AUMantagomir sdASO's concentration and on its affinity relative to those targets.

Mechanism

Mechanism of action

MicroRNA binding: AUMantagomir sdASOs are typically fully complementary (or heavily complementary) to the microRNA sequence of interest. A single microRNA (~22 nucleotides) can be entirely bound by the AUMantagomir sdASO. Upon binding, the ASO-microRNA duplex can prevent the microRNA from assembling with the RNA-induced silencing complex (RISC) or from attaching to its target mRNAs. This neutralizes the microRNA's regulatory effect.

MicroRNA inhibition vs. degradation: Depending on the design, an AUMantagomir sdASO can work by steric blocking (binding the microRNA and preventing it from functioning) and/or by recruiting RNase H1 to degrade the microRNA. In many cases an AUMantagomir sdASO results in the microRNA being degraded as part of an RNA:DNA hybrid, thereby reducing the microRNA's abundance.

Self-delivery: Like our other products, AUMantagomir sdASOs enter cells without needing a transfection reagent. This is crucial for microRNA studies because many cell types of interest (neurons, primary tissue cells, etc.) are difficult to transfect.

Three stages inside a cell, left to right: one uniform oligonucleotide strand with no gap pairs end to end with a short microRNA; the completed duplex is held under a warm glow while an Argonaute protein below it, a translucent two-lobed body with an open groove, stays empty; a messenger RNA runs from its capped start to its beaded tail through a ribosome, two rounded subunits joined on it, which reads it. Nothing is cut and no enzyme appears. The cell membrane stands at the left, the nucleus at the top right and a mitochondrion below. — Stage 1
AUMantagomir sdASOs enter cells without needing a transfection reagent.
Stage 2
AUMantagomir sdASOs are typically fully complementary (or heavily complementary) to the microRNA sequence of interest. A single microRNA (~22 nucleotides) can be entirely bound by the AUMantagomir sdASO.
Stage 3
Upon binding, the ASO-microRNA duplex can prevent the microRNA from assembling with the RNA-induced silencing complex (RISC) or from attaching to its target mRNAs. This neutralizes the microRNA's regulatory effect.

Applications

Applications and use cases

Functional studies of microRNAs
If you hypothesize a certain microRNA regulates a pathway (for example, miR-134-5p in synaptic plasticity or let-7a-5p in angiogenesis), inhibit the microRNA with AUMantagomir sdASO and read the outcome.
Target discovery
Use AUMantagomir sdASO to identify what genes a microRNA controls. After treatment with an AUMantagomir sdASO and transcriptomic analysis, the transcripts that rise are likely direct or indirect targets of the microRNA.
Disease models
Many diseases involve dysregulated microRNAs (fibrosis, cancer, neurodegeneration). AUMantagomir sdASOs are used in cellular and animal models of disease to model sustained microRNA inhibition.
Difficult-to-transfect cells
AUMantagomir sdASOs can enter primary macrophages and neurons in culture without a transfection reagent, electroporation, or a viral vector.

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

See where it works

Experimental considerations

The working concentration is higher than that of a transfected oligonucleotide. The recommended working range for AUMantagomir sdASOs is 5-20 μM. For AUMantagomir toASO, delivered with a transfection reagent, the recommended range is 50-100 nM.

Because an AUMantagomir sdASO binds the microRNA rather than always degrading it, a reduction in microRNA level is not always detected by qRT-PCR. De-repression of known target genes is the more reliable measure of inhibition.

In the literature