Preparation of AteloGene® Systemic Use | Koken Co., Ltd.

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Preparation of AteloGene® Systemic Use

Download the manual for AteloGene® Systemic Use 

Preparation instructions

In this section, the numbers ① – ⑦ refer to the ”Kit contents” on product page. Implement measures to secure an RNase-free environment, as far as possible, to avoid the degradation of siRNA/miRNA in advance.

1) Preparation of AteloGene®

Attach the ⑥18G needle to the ①AteloGene® prefilled syringe. Eject the whole amount (600 μL) into the ④Microtube. After ejection, cool the microtube containing “AteloGene®” on ice.

An excess amount of “AteloGene®” is provided in the ①Prefilled syringe so that 600 μL of AteloGene® can be injected into the ④Microtube regardless of losses during the preparation steps.

2) Preparation of siRNA/miRNA solution

Prepare 5-10 μM and 20-40 μM siRNA/miRNA solutions for local and systemic administrations, respectively.

a. Using siRNA/miRNA stock solutions
Adjust the concentration of the siRNA/miRNA solution with ②10×siRNA buffer and ③Sterilized water to a buffer concentration of 1×. Cool on ice.

b. Using lyophilized siRNA/miRNA
Prepare 1×siRNA buffer by diluting ②10×siRNA buffer with ③Sterilized water. Add 1×siRNA buffer to prepare siRNA/miRNA solutions of the desired concentrations. Cool on ice.

Although other buffers such as TE may be used for preparing the siRNA/miRNA solution, ②10×siRNA buffer is recommended for obtaining the best results.

3) Preparation of AteloGene® & siRNA/miRNA complex

While cooling on ice, gently pour 600 μL of the siRNA/miRNA solution onto 600 μL of AteloGene® in the ④Microtube. Slowly rotate and mix the solutions at 4℃ for 20 minutes. To avoid forming bubbles, the rotation speed should be approximately 4 r.p.m. (in the case of a rotator with a diameter of 20 cm).

4) Deformation of bubbles and preparation for administration

Centrifuge the mixture at 10,000 r.p.m. for 1 minute at 4℃ to deform bubbles. Attach the ⑥18G needle to the ⑤Disposable syringe and slowly draw the mixture while avoiding forming bubbles. Replace the needle with the ⑦26G injection needle and keep the syringe refrigerated until immediately before administration.