Invitrogen™ Invivofectamine™ 3.0 Reagent

Product Code.	No. of Reactions	unitSize
15331018	10	Each
15421144	50 Reactions	1 set

Product Code. 15331018 Supplier Invitrogen™ Supplier No. IVF3001

Description

An animal origin-free lipid nanoparticle designed for high efficiency in vivo delivery of siRNA and miRNA to mouse liver cells following tail vein injection

Using Invivofectamine™ 3.0 Reagent
Invitrogen™ Invivofectamine™ 3.0 Reagent is ideally suited for in vivo delivery of siRNA and miRNA duplexes. Invivofectamine™ 3.0/RNA complexes are delivered using a low-volume, low-pressure method with no toxicity and no stress response in the animal. Invivofectamine™ 3.0 can also be combined with BLOCK-iT™ fluorescent controls to track the biodistribution of the siRNA or miRNA.

For Research Use Only. Not for use in diagnostic procedures.

Specifications

• Classification: Animal Origin-Free
• Cell Type: Hepatocytes
• Sample Type: Synthetic siRNA, siRNA, miRNA, RNAi
• Transfection Technique: Lipid-based Transfection, Lipid Nanoparticles, LNPs
• Species: Small Animal Models
• For Use With (Application): Transfection
• No. of Reactions: 10
• Product Line: Invivofectamine
• Product Type: Reagent
• Quantity: 10 reactions
• RNAi Type: siRNA, miRNA
• Serum Compatible: Yes
• Shipping Condition: Dry Ice

Frequently Asked Questions (FAQs)

How do I isolate RNA from liver or other organs for my in vivo siRNA experiment?

Isolating high-quality RNA is critical to the overall success of in vivo experiments.

What rate should I use to inject complexed material for a systemic injection for my in vivo siRNA experiment?

Invivofectamine 3.0 Reagent is designed to be delivered slowly and evenly; a 200-µL injection should take between 10 to 20 seconds.

How important is sterile technique for my in vivo siRNA experiment?

Sterile technique is vital to successful completion of in vivo experiments. All materials need to be sterile before injecting the complexed siRNA into animals.

Can I modify the encapsulation protocol to fit my schedule for my in vivo siRNA experiment?

The protocol is robust but specific. Protocol modifications will likely result in less than optimal performance.

What controls should I use for my in vivo siRNA experiment?

Both positive and negative controls are recommended when performing any siRNA experiment. We typically use Invitrogen in vivo siRNA against Factor VII with Invivofectamine 3.0 Reagent, as a positive control for experiments. It is also good practice to use PBS solution as a negative control.

How many mice should I inject with siRNA per treatment?

We recommend injecting 3-5 mice per treatment to assess the reproducibility of the results within an experiment.

Do I need to weigh my mouse/animal to determine in vivo siRNA dosing, or can I use the standard 200 µL of complexed material for systemic intravenous or intraperitoneal injections?

To ensure the best results and minimal toxicity, it is very important to determine the weight of each animal to provide the correct dose and to maintain uniform dosing of multiple animals. The recommended dose for mice is 1 mg/kg ( IV) to start and reduce the dose depending on the potency of your siRNA.

Should I use any particular strain of mouse in my in vivo siRNA experiments?

We recommend the use of inbred strains for in vivo experiments, because these strains provide a uniform experimental background in which to determine gene knockdown and downstream effects. In addition, we recommend purchasing animals that are in good health from a qualified vendor. Starting with sick or lethargic mice is likely to result in problems during the experiment

Do I need to test my siRNA in vitro before using it in vivo?

Yes! We recommend that you test at least 3 different siRNA duplexes in vitro at several different concentrations and use the siRNA duplex with the highest potency for your in vivo experiments. When moving from in vitro to in vivo experiments, it is also important to double check the species specificity of the duplex.

Do I need to concentrate my siRNA before complexing with Invivofectamine 3.0 Reagent?

Your siRNA must be at least 1.2 mg/mL in RNase/DNase-free water before complexing. siRNA stock concentrations lower than 1.2 mg/mL will result in low encapsulation efficiencies. We also recommend that you confirm the siRNA concentration before complexing with Invivofectamine 3.0 Reagent to verify the correct concentration. After resuspension in RNase/DNase-free water, measure the OD at 260 nm of several serial dilutions of the stock and calculate the concentration.

What level of siRNA purity do you recommend for in vivo experiments?

We recommend that your siRNA is of the highest quality available. In vivo purity siRNA from Invitrogen or highly purified and desalted siRNA is required for good results. Failures in synthesis, excess salts, free nucleotides, and improperly purified siRNA may result in poor performance in vivo.

The Invivofectamine 3.0 Reagent (Cat. No. IVF3001) product page says the product is for 10 reactions. Does this mean it is for 10 siRNA?

No, "10 reactions" means it is for 10 injections into mice.

Can I use Invivofectamine 3.0 Reagent for transfection of mRNA?

Sorry, we have not tested this. We only have validated this reagent in-house for vivo delivery of siRNA and miRNA to the mouse liver by tail vein injection.

For mRNA in vivo delivery, please check out our Vivofectamine delivery solutions.

What is the highest amount of siRNA that can be complexed with Invivofectamine 3.0 Transfection Reagent?

Keep the siRNA concentration at 2.4 mg/mL and follow protocol instructions for complex preparation, keeping the siRNA to Invivofectamine 3.0 Transfection Reagent ratio at 1.2:1 (µg:µL).

What is the difference between Invivofectamine 2.0 Reagent and Invivofectamine 3.0 Reagent?

Invivofectamine 3.0 Reagent is an improved formulation with the following increased benefits:

- Lower toxicity
- Higher transfection efficiency with less siRNA; sustained knockdown up to 2 weeks
- Increased stability
- Shorter overall workflow; no dialysis or diafiltration needed

I need to inject higher quantity of siRNA than what is recommended in the protocol. How would I do this?

The Invivofectamine 3.0 Reagent-siRNA complexes may be used without the 6-fold dilution, in concentrated form. However, the complexes should be either dialyzed or diafiltrated before use.

How long can I store my siRNA complexed with Invivofectamine 3.0 Reagent?

Invivofectamine 3.0 Reagent-siRNA complexes may be stored up to two weeks at 4°C.

Can I use Invivofectamine 3.0 Transfection Reagent to deliver siRNA to organs other than the liver?

The Invivofectamine 3.0 Reagent has been validated in-house only for in vivo delivery of siRNA and miRNA to the mouse liver by tail vein injection.

How many injections can I perform with the Invitrogen Invivofectamine 3.0 Starter kit?

You may perform up to 10 injections of 200 µL each of a 0.1 mg/mL siRNA complex solution (e.g., 20 g mouse, 1 mg (siRNA)/kg (body mass)).

For Research Use Only. Not for use in diagnostic procedures.