mRNA体内高效转染试剂是一种基于多肽的高效率低毒性mRNA转染试剂，可用于体外转染，对Hela、CHO、DC2.4、Vero、4T1等多种细胞系实现高转染效率；也可用于体内转染，在无肿瘤小鼠的肺部和多种肿瘤模型小鼠的肿瘤部位实现高选择性高效率的mRNA表达。产品以无菌的液体形式提供。
通常情况下，使用推荐用量，每毫升本试剂大约可以转染6孔板40个孔、24孔板200个孔、96孔板1000个孔、13只小鼠（体重20g）。
组分与储存：
| 组分货号 | 组分名称 | 物理外观 | 100μL | 500μL | 1.5mL | 储存 |
| R751610A | mRNA体内高效转染试剂 | 液体 | 100μL | 500μL | 1.5mL | 2-8℃ |
| R751610B | Transfection Buffer | 液体 | 1mL | 5mL | 15mL | 2-8℃ |
实验材料：
• 本产品，包含mRNA体内高效转染试剂和Transfection Buffer两种试剂
• ⾼纯度的核酸储液（0.5∼5 μg/μL）
• ⽆⾎清培养基（如 O778393 Optimal-MEM减血清培养基 (含酚红) ）
质量控制：
• 4 °C保存条件下有效期一年
• 本产品经严格的质量检验证明，⽆⽣物污染
注意事项：
•常温下制备核酸-mRNA体内高效转染试剂复合物，使用灭酶灭菌管，孵育时间20～30min。
•本产品应该在4°C密闭保存，避免反复冻融。
•体外转染时，初次使用应该优化核酸浓度与mRNA体内高效转染试剂的比例以得到最高的转染效率，测试使用推荐的两种产品用量。
•体外转染时，请使用适当保存和经常传代的健康细胞，并确保培养基无细菌、真菌或支原体污染，如细胞是近期复苏的液氮冻存细胞，请在转染前至少传代两次。
•体内转染时，可通过改变肿瘤体积的大小、mRNA的用量和浓度以及mRNA体内高效转染试剂用量对体内转染进行优化。
体外多细胞系转染GFP mRNA效果示例：
体内转染Firefly Luciferase mRNA效果示例：
体外转染时按照下表转染细胞。使用指定量的mRNA试剂以及对应的两种体积的mRNA体内高效转染试剂(优化时)。每种反应混合物体积为单个孔的体积，且考虑了移液差异。按比例计算其他孔的体积。
体内转染时，根据小鼠体重优化转染条件。mRNA体内高效转染试剂(μL)：mRNA(μg)可在3：1到7：1间调整。保证肿瘤体系在50mm³~200mm³最佳
表1.不同体重鼠的转染用量
鼠体重
| 建议瘤体积
| mRNA体内高效转染试剂用量
| mRNA用量
|
15 g
| 100 mm³
| 60μL
| 12 μg
|
20 g
| 100 mm³
| 75 μL
| 15 μg
|
25 g
| 100 mm³
| 95 μL
| 19 μg
|
30 g
| 100 mm³
| 115μL
| 23 μg |
The High-Efficiency In Vivo mRNA Transfection Reagent is a polypeptide-based mRNA transfection reagent with high efficiency and low toxicity. It can be used for in vitro transfection, achieving high transfection efficiency in various cell lines such as HeLa, CHO, DC2.4, Vero, and 4T1. It can also be used for in vivo transfection, enabling highly selective and efficient mRNA expression in the lungs of tumor-free mice and in tumor sites of various tumor model mice. The product is supplied as a sterile liquid.
Under normal circumstances, with the recommended dosage, each milliliter of this reagent can approximately transfect 40 wells of 6-well plates, 200 wells of 24-well plates, 1000 wells of 96-well plates, or 13 mice (with a body weight of 20 g).
Contents & Storage:
Component No.
| Component Name
| Appearance
| 100μL | 500μL | 1.5mL | Storage
|
| R751610A | High-Efficiency In Vivo mRNA Transfection Reagent
| Liquid
| 100μL | 500μL | 1.5mL | 2-8℃ |
| R751610B | Transfection Buffer | Liquid
| 1mL | 5mL | 15mL | 2-8℃ |
Experimental Materials:
• This product contains two reagents: High-Efficiency In Vivo mRNA Transfection Reagent and Transfection Buffer.
• High-purity nucleic acid stock solution (0.5–5 μg/μL)
• Serum-free medium (e.g., O778393 Optimal-MEM Reduced Serum Medium (with Phenol Red))
Quality Control:
• The shelf life is 1 year when stored at 4 °C.
• Strict quality inspection has confirmed that this product is free from biological contamination.
Precautions:
• Prepare the nucleic acid-High-Efficiency In Vivo mRNA Transfection Reagent complex at room temperature using enzyme-free and sterile tubes, with an incubation time of 20–30 minutes.
• This product should be stored tightly sealed at 4°C, and repeated freezing and thawing should be avoided.
• For in vitro transfection, when using it for the first time, the ratio of nucleic acid concentration to High-Efficiency In Vivo mRNA Transfection Reagent should be optimized to achieve the highest transfection efficiency, and the recommended dosages of the two products should be used for testing.
• For in vitro transfection, use healthy cells that are properly preserved and regularly passaged, and ensure that the culture medium is free from bacterial, fungal, or mycoplasma contamination. If the cells are recently revived from liquid nitrogen-frozen stocks, they should be passaged at least twice before transfection.
• For in vivo transfection, the in vivo transfection can be optimized by adjusting the tumor volume, the dosage and concentration of mRNA, and the dosage of High-Efficiency In Vivo mRNA Transfection Reagent.
Example of GFP mRNA Transfection Effect in Multiple Cell Lines In Vitro:
Example of In Vivo Firefly Luciferase mRNA Transfection Effect:
For in vitro transfection, transfect cells according to the table below. Use the specified amount of mRNA reagent and the corresponding two volumes of High-Efficiency In Vivo mRNA Transfection Reagent (during optimization). The volume of each reaction mixture refers to the volume for a single well, with pipetting variations taken into account. Calculate the volumes for other wells in proportion.
For in vivo transfection, optimize the transfection conditions according to the body weight of mice. The ratio of High-Efficiency In Vivo mRNA Transfection Reagent (μL) to mRNA (μg) can be adjusted within the range of 3:1 to 7:1. It is optimal to keep the tumor volume within 50 mm³ - 200 mm³.
Table 1. Transfection Dosages for Mice of Different Body Weights
Mouse Weight
| Recommended Tumor Volume
| Dosage of High-Efficiency In Vivo mRNA Transfection Reagent
| Dosage of mRNA
|
15 g
| 100 mm³
| 60μL
| 12 μg
|
20 g
| 100 mm³
| 75 μL
| 15 μg
|
25 g
| 100 mm³
| 95 μL
| 19 μg
|
30 g
| 100 mm³
| 115μL
| 23 μg |
危险品化学品经营许可证（带存储）