Product description

Fast LNP (in vivo) is a ready-to-use Lipid Nanoparticle (LNP) for in vivo transfection of mRNA in experimental animals. LNP can be encapsulated in one step without the need of specific encapsulation equipment, without the need of incubation, and the transfection effect is comparable to that of LNP encapsulated by professional encapsulation equipment, and it can meet a variety of drug delivery modes, including but not limited to intramuscular, intravenous, percutaneous, and intraperitoneal administration routes.

Transportation and storage

Transportation: temperature-controlled (2°C-25°C) transport, freezing prohibited.

Storage: Fluc mRNA Positive Control: -25°C to -15°C; other components: 2°C-8°C, freezing prohibited. Valid for 1 year.

Kit Components

Using protocol

Flowchart of in vivo delivery operation

Example: Injection of 30 μg mRNA with injection volume of 200 μL.

Recommended ratio: 1 μg mRNA : 1 μL Fast LNP Reagent.

1. Positive control preparation (Optional)

Take out the Fluc mRNA positive sample reagent tube, flick the wall of the tube with your finger, then place the tube in a centrifuge, centrifuge at 10000 rpm 30s and add 10 μL sterile nuclease-free water, flick the tube to mix well and centrifuge at 10000 rpm, 30s. Keep on ice for use.

2. mRNA premix preparation

Take 37.5 μL of mRNA (1 μg/μL, for example) in a sterile, nuclease-free tube, and add 175 μL of Fast LNP Buffer for mixing by pipette.

3. mRNA-LNP preparation

Add 37.5 μL of Fast LNP reagent to the above mRNA premix, pipette blow more than 10 times and mix well to obtain 250 μL of mRNA-LNP. Final mRNA concentration must be ≤0.2 μg/μL.

4. in vivo administration

200 μL of mixed mRNA-LNP was aspirated with a syringe and injected into mice according to a predetermined protocol.

5. Perform in vivo delivery efficiency assay in mice under the experimental endpoint conditions.

Example: Injection of 20 μg siRNA with injection volume of 100 μL.

Recommended ratio: 2 μg siRNA : 1 μL Fast LNP Reagent.

1. siRNA sample preparation

Resuspend siRNA by flicking the tube. Centrifuge at 10,000 rpm for 30 seconds. Add nuclease-free water to achieve ≥2 μg/μL siRNA concentration. Mix and centrifuge again (10,000 rpm, 30 seconds). Keep on ice.

2. siRNA premix preparation

Transfer 10 μL siRNA (2 μg/μL, for example) into a sterile nuclease-free tube. Add 80 μL Fast LNP Buffer and mix.

3. siRNA-LNP Preparation

Add 10 μL Fast LNP Reagent to the premix. Pipette vigorously (>10 times) to homogenize. siRNA-LNP volume: 100 μL. Final siRNA concentration must be ≤1 μg/μL.

4. in vivo administration

100 μL of mixed siRNA-LNP was aspirated with a syringe and injected into mice according to a predetermined protocol.

5. Perform in vivo delivery efficiency assay in mice under the experimental endpoint conditions.

Dosage Conversion for Each Component

(1) The dosage of Fast LNP Buffer generally depends on the target injection volume. Fast LNP Buffer volume = injection volume - RNA volume - Fast LNP reagent.

(2) The recommended dosage of each component for different dosage regimens is shown in Table 1.

(3) The recommended dosages for different delivery modes are shown in Table 2.

Precautions

1. Do not freeze or thaw Fast LNP reagent and RNA-LNP.

2. The configured mRNA-LNP can be directly administered to animals without room temperature incubation.

3. For your safety and health, please wear a lab coat and disposable gloves.

4. This product is intended for scientific use only and should not be used in humans.

Table 1. The recommended dosage of each component for different dosage regimens

Table 2. The recommended dosages for different delivery modes