LNP RNA Transfection Reagent (Common Cells) is a novel ready-to-use transfection reagent developed based on lipid nanoparticle (LNP) technology. Leveraging the unique transmembrane transport and endosomal escape mechanisms of LNPs, it enables efficient delivery of RNA and is widely applicable for transfecting various types of RNA into adherent or suspension conventional cell lines.
User-Friendly: No specialized encapsulation equipment required. RNA-LNP transient encapsulation is completed via one-step mixing with no incubation period; transfection can be performed immediately after preparation, and medium replacement is unnecessary post-transfection.
High Efficiency: The encapsulation efficiency of trace samples can exceed 80%, supporting robust transfection performance across common cell lines.
Low Cytotoxicity & Safety: This product is ethanol-free and formulated with novel biodegradable lipid materials, resulting in extremely low cytotoxicity. In addition, the reagent is sterile-filtered through a 0.2 μm microporous membrane.
Components List
| L1375584 | Components | Appearance | 1.5 ml | 3.0 ml | 5.0 ml | Storage | | L1375584A | LNP RNA Transfection Reagent (Common Cells) | Liquid | 1.5 ml | 3.0 ml | 5.0 ml | 2-8℃ | | L1375584B | Buffer | Liquid | 8mL | 2×8mL | 3×8mL | 2-8℃ |
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Transportation and Storage
Ship under controlled ambient temperature (≤25°C). Store at 2°C ~ 8°C. Do NOT freeze!
Scope of Application
This LNP RNA Transfection Reagent (Common Cells) is suitable for in vitro RNA transfection of the vast majority of eukaryotic cells. Applicable RNA types include siRNA, miRNA, gRNA, mRNA, saRNA, circRNA, etc.This product is for research use only.
Precautions
(1) Nuclease-free water is the optimal solvent for stock RNA solutions. The concentration of mRNA shall be no less than 200 ng/μL; short-chain RNA no less than 100 ng/μL; siRNA no less than 10 μM. Other solvents may compromise transfection efficiency. Please contact us for relevant details.
(2) Prepare RNA premix by diluting stock RNA with the dedicated Buffer supplied in this kit. Do not use PBS or cell culture medium.
(3) Adjust the final concentration of RNA premix to 40 ng/μL. For co-transfection of multiple RNAs (e.g. CRISPR Cas9 mRNA and gRNA), the concentration refers to the total concentration of Cas9 mRNA plus gRNA. The recommended mass ratio of Cas9 mRNA to gRNA is 1:1 ~ 1:2 (μg mRNA : μg gRNA).
(4) The RNA/LNP ratio is a critical parameter for LNP encapsulation. If adjusting the dosage per well, follow the instructions below:
1. For mRNA transfection and mRNA+gRNA co-transfection: if increasing or decreasing the mRNA dosage per well, proportionally adjust the volume of RNA-LNP complex added to each well.
2. For siRNA transfection: if increasing the siRNA dosage per well, proportionally increase the volume of RNA-LNP complex added to each well.
If reducing the siRNA dosage per well, the minimum volume of LNP transfection reagent shall not be lower than Recommended Dosage II. Take a 96-well plate as an example: when siRNA dosage is reduced to 0.5 pmol per well, adjust the siRNA premix concentration to 0.5 μM with a minimum reagent volume of 0.5 μL per well; all other steps remain unchanged.
(5) RNA-LNP complexes remain stable at room temperature for up to 8 hours. Do not freeze RNA-LNP complexes.
(6) Medium replacement is not required before or after transfection. Complete medium containing serum and antibiotics shows no significant interference with transfection results. Medium exchange or routine cell maintenance can be performed 6 hours post-transfection.
Appendix A: Cell Transfection Protocol
1. Operating Procedure for LNP RNA Transfection Reagent (Common Cells) (Recommended Dosage I)
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2. Reagent Dosage Per Well for Different Culture Vessels (Recommended Dosage I)
| RNA Cell Transfection Steps | Reagents | 96-well plate | 24-well plate | 6-well plate |
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| 1) Preparation of 40 ng/μL RNA Premix | Buffer (stock RNA concentration: 1 μg/μL for reference) | 2.4 μL | 12 μL | 60 μL |
| RNA | 0.1 μg | 0.5 μg | 2.5 μg | | 2) Preparation of RNA-LNP Complex | LNP RNA Transfection Reagent (Common Cells) | 2.5 μL | 12.5 μL | 62.5 μL |
| RNA Premix | 2.5 μL | 12.5 μL | 62.5 μL | | 3) Addition to Cells | RNA-LNP Complex | 5 μL | 25 μL | 125 μL |
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| siRNA Cell Transfection Steps | Reagents | 96-well plate | 24-well plate | 6-well plate |
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| 1) Preparation of 1 pmol/μL siRNA Premix (equivalent to 1 μM) | Buffer (stock siRNA concentration: 10 μM for reference) | 0.9 μL | 4.5 μL | 22.5 μL |
| siRNA | 1 pmol | 5 pmol | 25 pmol | | 2) Preparation of siRNA-LNP Complex | LNP RNA Transfection Reagent (Common Cells) | 1 μL | 5 μL | 25 μL |
| siRNA Premix | 1 μL | 5 μL | 25 μL | | 3) Addition to Cells | siRNA-LNP Complex | 2 μL | 10 μL | 50 μL |
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3. Operating Protocol for LNP RNA Transfection Reagent (Common Cells) (Recommended Dosage II)
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4. Reagent Dosage Per Well for Different Culture Vessels (Recommended Dosage II)
| RNA Cell Transfection Steps | Reagents | 96-well plate | 24-well plate | 6-well plate |
|---|
| 1) Preparation of 40 ng/μL RNA Premix | Buffer (stock RNA concentration: 1 μg/μL for reference) | 2.4 μL | 12 μL | 60 μL |
| RNA | 0.1 μg | 0.5 μg | 2.5 μg | | 2) Preparation of LNP Premix | Buffer | 1.25 μL | 6.25 μL | 31.25 μL |
| LNP RNA Transfection Reagent (Common Cells) | 1.25 μL | 6.25 μL | 31.25 μL | | 3) Preparation of RNA-LNP Complex | LNP Premix | 2.5 μL | 12.5 μL | 62.5 μL |
| RNA Premix | 2.5 μL | 12.5 μL | 62.5 μL | | 4) Addition to Cells | RNA-LNP Complex | 5 μL | 25 μL | 125 μL |
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| siRNA Cell Transfection Steps | Reagents | 96-well plate | 24-well plate | 6-well plate |
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| 1) Preparation of 1 pmol/μL siRNA Premix (equivalent to 1 μM) | Buffer (stock siRNA concentration: 10 μM for reference) | 0.9 μL | 4.5 μL | 22.5 μL |
| siRNA | 1 pmol | 5 pmol | 25 pmol | | 2) Preparation of LNP Premix | Buffer | 0.5 μL | 2.5 μL | 12.5 μL |
| LNP RNA Transfection Reagent (Common Cells) | 0.5 μL* | 2.5 μL* | 12.5 μL* | | 3) Preparation of siRNA-LNP Complex | LNP Premix | 1 μL | 5 μL | 25 μL |
| siRNA Premix | 1 μL | 5 μL | 25 μL | | 4) Addition to Cells | siRNA-LNP Complex | 2 μL | 10 μL | 50 μL |
* The indicated volume of LNP RNA Transfection Reagent (Common Cells) per well is the minimum dosage for siRNA transfection. |
Appendix B: Recommended Co-transfection Protocol for CRISPR/Cas9 mRNA + gRNA
1. Operating Procedure for Co-transfection of Cas9 mRNA and gRNA
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2. Reagent Dosage Per Well for Different Culture Vessels
| Cas9 mRNA + gRNA Cell Transfection Steps | Reagents | 96-well plate | 24-well plate | 6-well plate |
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| 1) Preparation of 40 ng/μL RNA Premix | Buffer | 4.8 μL | 24 μL | 120 μL |
| Cas9 mRNA (stock mRNA concentration: 1 μg/μL for reference) | 0.1 μg | 0.5 μg | 2.5 μg |
| gRNA (stock gRNA concentration: 1 μg/μL for reference) | 0.1 μg | 0.5 μg | 2.5 μg | | 2) Preparation of LNP Premix | Buffer | 2.5 μL | 12.5 μL | 62.5 μL |
| LNP RNA Transfection Reagent (Common Cells) | 2.5 μL | 12.5 μL | 62.5 μL | | 3) Preparation of RNA-LNP Complex | LNP Premix | 5 μL | 25 μL | 125 μL |
| RNA Premix | 5 μL | 25 μL | 125 μL | | 4) Addition to Cells | RNA-LNP Complex | 10 μL | 50 μL | 250 μL |
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