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BioShuttle siRNA/miRNA in vivo transfection reagent (in vivo)

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GoodsCode: KX0110050
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  • 0.8ml
    300.00
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Product Informations

  • Goods Code KX0110050
  • Goods Name BioShuttle siRNA/miRNA in vivo transfection reagent (in vivo)
  • Category 525
  • Shipping and Storage Conditions Transported at room temperature, stored at 4°C, stable storage period of 12 months. long-term storage at -20°C;
  • Precautions 注意事项
    1. 使用高纯度的RNA是转染成功的关键因素，在转染前确定RNA的含量和纯度（OD260/280＞1.8），实验过程中使用DEPC处理过的耗材和试剂，避免RNase污染；另外RNA制备时盐含量高会导致复合物形成时产生沉淀，因此应将RNA重悬于5%葡萄糖溶液（由DEPC水配制）或DEPC水中；
    2. 体内转染所用RNA以及稀释液均为无菌制品，且操作步骤全程无菌；
    3. 体内转染时按需现配现用；
    4. 为了达到更好的转染效果，可根据实验需要优化RNA与BioShuttle siRNA/miRNA体内转染试剂(in vivo)的配比，推荐优化比例范围为1:1~1:4 (w/v, RNA: BioShuttle siRNA/miRNA体内转染试剂(in vivo))；
    5. 本产品仅用于科学研究，不用于临床诊断和治疗。
  • Instruction Manual /files/uploads/instructions/2024/202403/KX0110050_BioShuttlesiRNAmiRNA体内转染试剂(in vivo)_说明书240314.pdf

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Product Details

Kit components Specification Save
BioShuttle siRNA/miRNA Transfection Reagent in vivo 0.8ml Store at 4℃
10% glucose solution (gift) * 4ml Store at 4℃
* Note: Here, a 10% glucose solution (w/v, DEPC water configuration) is provided as a free gift. If a larger quantity is required, sterile DEPC water can be used for self-preparation.

Product Description
BioShuttle siRNA/miRNA in vivo transfection reagent (in vivo) is a new type of highly efficient cationic polymer that can interact with RNA (including siRNA, miRNA, and recombinant RNA) to form nanocomplexes, thereby being used for in vivo delivery of RNA. BioShuttle siRNA/miRNA in vivo transfection reagent (in vivo) has the characteristics of high delivery efficiency and good safety. After BioShuttle siRNA/miRNA transfection reagent (in vivo) enters the cell through endocytosis, it expresses the characteristic of "proton sponge effect", buffers the pH value of the endosome, protects RNA from degradation, and the continuous proton influx can also cause endosome osmotic swelling and rupture, providing a mechanism for RNA to escape into the cytoplasm.

Product Features
High delivery efficiency
Good safety

Operation steps
（1）Reagent requirements
in order to form a BioShuttle siRNA/miRNA in vivo transfection reagent (in vivo)/RNA nanocomplex with uniform size and stable morphology, it is recommended to use a 5% sterile enzyme-free isotonic glucose solution (provide a 10% sterile enzyme-free glucose solution, and dilute the final concentration to 5% at in vivo injection);
If the salt content of the solution diluting RNA is high, it may cause precipitation when forming complexes with the BioShuttle siRNA/miRNA in vivo transfection reagent.
RNA was purified using PAGE or HPLC.
(2) Recommended RNA volume and injection volume
The RNA dosage was determined based on the animal model, administration route and target organ of action. Table 1 provides the recommended proportioned doses for RNA transfection in rodents.
The final RNA concentration of the injection should not exceed 0.5 μg/μL to avoid precipitation
The recommended usage ratio of RNA to BioShuttle siRNA/miRNA in vivo transfection reagent (in vivo) is 1:1 by mass volume (w/v, 1 μg RNA: 1 μL BioShuttle siRNA/miRNA in vivo transfection reagent (in vivo)).

Table 1. Recommended proportioned doses for RNA transfection in rodents
Animal Injection method Compound formula Optimal range of RNA Optimal range of injection volume
Mouse Intravenous injection 20 μgRNA+20 μL transfection reagent + glucose solution to 100 μL (the final concentration of the glucose solution is 5%) 20-60 μg
(1-3mg/kg)		 100-300 μL
 
Intraperitoneal injection 100 μgRNA+100 μL transfection reagent + glucose solution to 0.4 mL(the final concentration of the glucose solution is 5%) 100-200 μg
(4-8mg/kg)		 < 1 mL
Subcutaneous injection 5 μg RNA+5 μL transfection reagent + glucose solution to 10 μL(the final concentration of the glucose solution is 5%) 3-5 μg 5-15 μL
In situ injection 5 μg RNA+5 μL transfection reagent + glucose solution to 10 μL(the final concentration of the glucose solution is 5%) 5-10 μg
(0.25-0.5mg/kg)		 10-20 μL
Note: For other animals and in vivo transfection ratio doses of different injection methods, please contact technical support.
(3) Instructions for Use
a. Dilute the RNA stock solution with 10% glucose solution (usually prepared with sterile DEPC water) and mix well. The volume of the diluted RNA solution is half of the final required injection volume, and the final concentration of the glucose solution is 5%.
b. Dilute the BioShuttle siRNA/miRNA transfection reagent in vivo with 10% glucose solution. The volume of the required BioShuttle siRNA/miRNA in vivo transfection reagent (in vivo) is calculated based on the volume-to-mass ratio of transfection reagent to RNA at 1:1. The volume of the transfection reagent diluted should be half of the final injection volume, and the final concentration of the glucose solution should be 5%. Mix well.
c. Add the diluted BioShuttle siRNA/miRNA in vivo transfection reagent (in vivo) to the diluted RNA and gently mix well;
d. Incubate at room temperature for 15 minutes. The complex formed at this time can be stably stored at room temperature for 2 hours.
e. Monitor gene expression at an appropriate time (e.g., 6 to 96 hours after injection), with the specific time depending on the injection method and the target organ.
f. Take intravenous injection in mice as an example to illustrate the preparation method of the complex. For example, the experimental injection dose was 25 μg RNA per mouse, and the concentration of the RNA stock solution was 1 μg/μL. The injection volume is 100 μL per mouse. The recommended usage ratio of RNA to BioShuttle siRNA/miRNA in vivo transfection reagent (in vivo) is 1:1 by mass volume (w/v, 1 μg RNA: 1 μL BioShuttle siRNA/miRNA in vivo transfection reagent (in vivo)). Therefore, the following solution needs to be prepared for a single mouse:
i. RNA working solution, dilute the RNA stock solution with 10% glucose solution; Take 25 μL of RNA stock solution, add 25 μL of 10% glucose solution, and mix well.
ii. Working solution of BioShuttle siRNA/miRNA in vivo Transfection Reagent (in vivo), dilute BioShuttle siRNA/miRNA in vivo transfection reagent (in vivo) with 10% glucose solution; Take 25 μL of BioShuttle siRNA/miRNA transfection reagent (in vivo), add 25 μL of 10% glucose solution, and mix well. After the RNA and BioShuttle siRNA/miRNA in vivo transfection reagent (in vivo) working solution were prepared, add the BioShuttle siRNA/miRNA in vivo transfection reagent (in vivo) working solution to the RNA working solution and gently mix well; Incubate at room temperature for 15 minutes and then inject through the tail vein.
The preparation processes of the other administration methods are the same as those in this example (Table 2)
Table 2. Examples of proportioned doses for RNA transfection in vivo
Injection method (injection volume) Injection volume RNA stock solution (1 μg/μL) A 10% glucose solution for diluting the RNA stock BioShuttle siRNA/miRNA Transfection Reagent in vivo Dilute the volume of 10% glucose solution of BioShuttle siRNA/miRNA in vivo transfection reagent
Intravenous injection (25 μg RNA per piece) 100 μL 25 μL 25 μL 25 μL 25 μL
Intraperitoneal injection (50 μg RNA per piece) 200 μL 50 μL 50 μL 50 μL 50 μL
Subcutaneous injection (5 μg RNA per piece) 20 μL 5 μL 5 μL 5 μL 5 μL
In situ injection (10 μg RNA per piece) 40 μL 10 μL 10 μL 10 μL 10 μL

Note: The concentration of the RNA stock solution is 1 μg/μL

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Product Q&A

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Q1: Does this in vivo transfection reagent have bone tissue targeting?

Q2: Must the RNA used be purified by PAGE and HPLC? Are there other purification methods for siRNA and miRNA?

Q3: Must the RNA used be purified by PAGE and HPLC methods? Are there any other purification methods for siRNA and miRNA?

Q4: Are the purification methods for siRNA and miRNA different from mRNA purification?

Q5: Are the purification methods of siRNA and miRNA different from those of mRNA?

Q6: Can the RNA and transfection reagent complex be diluted with other reagents?

Q7: Can the RNA and transfection reagent complex be diluted with other reagents?

Q8: Why is glucose solution needed for dilution? To reduce injection volume, can the glucose diluent be omitted?

Q9: May I ask why glucose solution is needed for dilution? Can glucose diluent be omitted to reduce the injection volume?

Q10: Can both RNA transfection reagents be used for RNA and DNA co-transfection?

Q11: What is the difference between the [RNA In Vitro Transfection Reagent] and the [RNA In Vivo Transfection Reagent]?

Q12: What are the differences between "RNA in vitro transfection reagents" and "RNA in vivo transfection reagents"?

Q13: Is your transfection reagent GMP grade?

Q14: For the siRNA in vivo transfection reagent, the product Q&A mentions non-targeting, most obvious in lung and liver. The customer's target organ is the spleen. Do you have relevant data for this?

Q15: How to verify efficiency?

Q16: What factors affect knockdown efficiency?

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