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miRNA mimic oligos

AUMmimic sdASO

Self-delivering antisense oligonucleotides for microRNA replacement

Species

Nothing is chosen: a page and an order name one species.

The mature name, without the species prefix.

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A self-delivering microRNA mimic, one uniform strand with no gap, is taken up by endocytosis: 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 strand is held inside an Argonaute protein, a translucent two-lobed body, that sits on a messenger RNA near its beaded tail, the strand paired with a short stretch of the message under a warm glow. The messenger RNA runs through the complex unbroken, from its capped start to its tail; nothing is cut, no enzyme appears, and a mitochondrion lies below.

AUMmimic sdASOs are self-delivering antisense oligonucleotides (ASOs) that supply a chosen mature microRNA (miRNA). MicroRNAs are small regulatory RNAs that typically bind to mRNAs and repress their translation or cause degradation. Where AUMantagomir sdASO inhibits a mature microRNA, AUMmimic sdASO stands in for one: the same self-delivering family, the opposite action.

Many of the primary cells and disease-relevant models in which a microRNA is replaced 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, AUMmimic sdASOs are self-delivering. They enter the cell without a transfection reagent, electroporation, or a viral vector. Add them to the culture medium, or inject them in vivo. Our AUMsilence platform uses chemical modifications that enable self-delivery.

  • No transfection required

    As with every self-delivering product, no transfection reagents, electroporation, or viral vectors are needed: add AUMmimic sdASO to the culture medium, or inject it in an animal model with no nanoparticle or viral delivery.

  • The counterpart of AUMantagomir sdASO

    AUMantagomir sdASO inhibits a mature microRNA; AUMmimic sdASO supplies one.

  • One mimic per mature microRNA

    The two arms of a hairpin are different microRNAs, so a mimic is made against one of them and is named with its arm: miR-21-5p or miR-21-3p, never miR-21.

  • Two purifications, one labeled version

    Which purification you need is set by the study: RPC for in vitro, cellular studies, and HPLC for in vivo, animal studies.

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

One designed sequence per mature microRNA. The scramble, positive and labeled scramble controls of the self-delivering class are offered on the order page.

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

At a glance

Class
Self-delivering antisense (sdASO)
Targets
miRNA mimic
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

No protocol page is written for this product yet: the steps below are the AUMsilence sdASO protocol's, the self-delivering class's, in its own words. The full protocol carries the amounts per plate format, the tips and the troubleshooting.

Read the AUMsilence sdASO protocol

  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 treatment is typical.

  2. Step 2

    AUMsilence sdASO stock preparation

    Prepare AUMsilence sdASO 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

    AUMsilence sdASO delivery to cells

    Add AUMsilence sdASO to the cells at the desired final concentration. The recommended working range is 5-20 μM, with a starting concentration of 10 μM.

  4. Step 4

    Incubation and analysis

    Incubate cells with AUMsilence sdASO and analyze knockdown at appropriate time points. Knockdown is typically observed 24 to 72 hours after treatment, depending on the cell type and the target gene. Longer time points may be required in some systems. Protein knockdown can also be observed from 24 hours after treatment. Where the target protein is long-lived, existing protein must first turn over, so a later time point may be required.

FigureThe workflow in cell culture, in four steps

In vitro, the four steps

  1. Step 1

    Prepare cells

    Seed the cells in a 24-well plate; the oligonucleotide waits in solution.

  2. Step 2

    Add to the medium

    Add the self-delivering oligonucleotide straight to the medium: no transfection reagent, no carrier.

  3. Step 3

    Incubate

    Incubate at 37 degrees C in 5% carbon dioxide, humidified, for the exposure time.

  4. Step 4

    Quantify knockdown

    Measure the knockdown: RT-qPCR, immunoblot, flow cytometry.

    Schematic of the three readouts, not data.

For research use only. Not for use in diagnostic or therapeutic procedures.
Sources
  1. 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.
  2. 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 AUMmimic sdASO

  • No transfection required

    As with every self-delivering product, no transfection reagents, electroporation, or viral vectors are needed: add AUMmimic sdASO to the culture medium, or inject it in an animal model with no nanoparticle or viral delivery.

  • The counterpart of AUMantagomir sdASO

    AUMantagomir sdASO inhibits a mature microRNA; AUMmimic sdASO supplies one. Both are self-delivering, so the two directions of a microRNA experiment, inhibition and replacement, run in the same format.

  • One mimic per mature microRNA

    The two arms of a hairpin are different microRNAs, so a mimic is made against one of them and is named with its arm: miR-21-5p or miR-21-3p, never miR-21. This product is sold as a single designed sequence per target, in human, mouse or rat.

  • Two purifications, one labeled version

    Which purification you need is set by the study: RPC for in vitro, cellular studies, and HPLC for in vivo, animal studies. A fluorescently labeled version, AUMmimiclab sdASO, is ordered to monitor cellular uptake; the sizes and the dyes of both are listed with the order form.

Mechanism

Mechanism of action

Self-delivery: Like our other self-delivering products, AUMmimic sdASOs enter cells without a transfection reagent. Our AUMsilence platform uses chemical modifications that enable self-delivery.

Replacement: An AUMmimic sdASO stands in for the mature microRNA it is designed against. It supplies that microRNA's activity, so the microRNA's targets are regulated as the natural microRNA regulates them. AUMantagomir sdASO takes the opposite action, binding the mature microRNA and inhibiting it.

Design: The AUMsilence platform designs the sequence from the name it is given, and the name has to be exact, because the arm decides which sequence is made. The name is miR or let, the number and the arm, as in miR-21-5p. The species control supplies the species prefix, hsa, mmu or rno, so the name is written without it.

FigureHow AUMmimic sdASO acts
Three stages inside a cell, left to right: one uniform oligonucleotide strand with no gap slides into an Argonaute protein, a translucent two-lobed body; the loaded complex sits on a messenger RNA with the strand paired to a short stretch of it under a warm glow, the message running through unbroken from its capped start to its beaded tail; on a second message the loaded complex holds its place near the tail while a ribosome, two rounded subunits, is stalled at the capped start. Nothing is cut and no enzyme appears. The cell membrane stands at the left, the nucleus at the top right and a mitochondrion at the lower right.
  1. Stage 1

    Like our other self-delivering products, AUMmimic sdASOs enter cells without a transfection reagent.

  2. Stage 2

    An AUMmimic sdASO stands in for the mature microRNA it is designed against.

  3. Stage 3

    It supplies that microRNA's activity, so the microRNA's targets are regulated as the natural microRNA regulates them.

Applications

Applications and use cases

  • Restoring a microRNA

    Where a microRNA is lost or reduced in a model, supply it with AUMmimic sdASO and read the outcome: the replacement arm of a microRNA study.

  • Inhibition and replacement, side by side

    Pair AUMmimic sdASO with AUMantagomir sdASO against the same mature microRNA to read its function from both directions, in the same self-delivering format.

  • Difficult-to-transfect cells

    Self-delivering antisense oligonucleotides are used where transfection can fail (e.g., primary cells, neurons and immune cells). AUMmimic sdASO is added to the medium in the same way, with no delivery step.

  • In vivo

    HPLC purification is the grade for in vivo, animal studies. Self-delivering antisense oligonucleotides are dissolved in a buffer such as phosphate-buffered saline (PBS) or saline and injected, with no nanoparticle or viral delivery.

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 AUMmimic sdASOs is 5-20 μM.

At the bench

Controls and what arrives

Add to culture medium or inject in vivo. The controls and the How much to order page are written for this product.

View protocolsControls and validationHow 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 AUMmimic sdASO

Name the mature microRNA with its arm and choose the species; the AUMsilence platform designs the sequence, and the price is on the order page.

Request a quoteHow to citeView protocols

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