OptiLNP RNA Transfection Reagent (Stem Cells) 

Lipid nanoparticles (LNP) are a lipid-based nanoscale drug delivery system widely used for the delivery of nucleic acids (such as mRNA, siRNA, DNA, etc.) and small molecule drugs.

Naked RNA is a negatively charged hydrophilic macromolecule that faces challenges in entering cells due to electrostatic repulsion from the cell membrane and is easily degraded by RNases. By encapsulating RNA within LNP, RNA can effectively traverse the cell membrane and be released into the cytoplasm, enabling efficient delivery.

MCE OptiLNP RNA Transfection Reagent (Stem Cells) is a ready-to-use transfection reagent based on LNP technology. It utilizes the unique transmembrane transport and escape mechanism of LNP to achieve efficient delivery of RNA, making it suitable for transfection of various types of RNA in stem cells.

Features of MCE OptiLNP RNA Transfection Reagent (Stem Cells)

1. Lipid Nanoparticle (LNP) Technology: Utilizes LNP carriers to effectively protect RNA from degradation and immune system clearance during the delivery process.

2. Efficient Intracellular Delivery: The unique transmembrane transport and escape mechanisms of LNPs achieve an encapsulation rate of over 80% for small sample amounts, enabling efficient transfection of RNA in stem cells.

3. Wide Applicability: Suitable for in vitro RNA transfection of various types of stem cells. Supports a variety of RNA types such as siRNA, miRNA, gRNA, mRNA, saRNA, and circRNA.

4. User-Friendly Operation: No encapsulation equipment is required; RNA-LNP encapsulation and transfection can be completed in a single mixing step, and there is no need to replace with fresh culture medium after transfection.

5. Biocompatibility: Low cytotoxicity, effectively balancing high transfection efficiency with cell viability, and providing gentle action.

6. Safety: The formulation uses biodegradable lipid materials to ensure safety.

4℃, 1 year

Do not freeze.

1. This product has been filtered using a 0.22 μm membrane.

2. For optimal transfection efficiency, it is recommended to dissolve the RNA stock in nuclease-free sterile water. Other solvents may affect transfection efficiency.

3. Recommended RNA concentration: mRNA > 200 ng/μL, Short-chain RNA > 100 ng/μL, siRNA > 10 μM.

4. It is recommended to prepare RNA pre-mixes using the dilution buffer provided with the kit to dilute the RNA stock solution.

5. The recommended concentration for RNA premix is 40 ng/μL, with a final concentration of 20 ng/μL. For co-transfection of multiple RNAs, the RNA premix concentration should be the sum of the concentrations of all RNAs.

6. Common short-chain RNAs (e.g., siRNA) typically weigh between 13 - 15 μg per 1 nmol without special modifications. The exact weight can be calculated based on the specific nucleic acid sequence and modifications.

7. The RNA/LNP ratio is crucial for LNP encapsulation. If the per-well RNA amount needs adjustment, the volume of RNA-LNP complex added to each well should be proportionally adjusted.

8. RNA-LNP complexes can be stably stored at room temperature for up to 8 h. Do not store RNA-LNP complexes under freezing conditions.

9. There is no need to replace the culture medium before or after transfection. Complete medium containing serum and antibiotics does not significantly affect transfection efficiency.

10. 6 h after transfection, standard cell culture processes, such as medium replacement, can be performed.

11. Cell conditions greatly influence transfection efficiency. It is recommended to use cells in good growth condition for transfection.

12. There are many factors affecting transfection efficiency, such as cell type, cell state and density, nucleic acid quality and concentration, ratio of transfection reagents to nucleic acids, etc. It is recommended to perform pre-experiment to determine optimal transfecti

Take the 24-well plate as an example; the cell volume of other culture well plates can be designed according to the conventional experiments.

For adherent cells: Plate the cells digested with trypsin one day before transfection (0.1 - 1 × 10⁵ cells plated in a 24-well cluster plate), until the density reaches 30% - 40% for cell transfection.

For suspension cells: Plate the cells before transfection and suspend in fresh medium (0.5 - 2.5 × 10⁵ cells/500 μL medium).

Note: The viability and general health of cells prior to transfection significantly affect the transfection result. Cells should be at least 90% viable prior to transfection and have had sufficient time to recover from passaging.

1. Prepare RNA Pre-mix Solution: Ddilute the RNA stock solution with the dilution buffer. Adjust the mRNA concentration to 40 ng/μL or the siRNA concentration to 3 pmol/μL.

2. Prepare RNA-LNP Complex (Choose one of the following two methods):

a. No Dilution of Transfection Reagent: Refer to Tables 1 and 2. Mix equal volumes of transfection reagent and RNA pre-mix directly. Pipette up and down at least 20 times to ensure thorough mixing.

b. Dilution of Transfection Reagent: Refer to Tables 3 and 4. Dilute the transfection reagent 2-fold with the dilution buffer to prepare the transfection reagent pre-mix. Mix equal volumes of the transfection reagent pre-mix and RNA pre-mix. Pipette up and down at least 20 times to ensure thorough mixing.

Add RNA-LNP Complex to each well, mix gently and incubator for further culture.

After incubation of 24 - 48 h, the transfection effect can be analyzed by fluorescence detection, Western Blot, ELISA, flow cytometry, reporter gene, immunofluorescence staining and other methods according to the experimental needs.

1. This product has been filtered using a 0.22 μm membrane.

2. For optimal transfection efficiency, it is recommended to dissolve the RNA stock in nuclease-free sterile water. Other solvents may affect transfection efficiency.

3. Recommended RNA concentration: mRNA > 200 ng/μL, Short-chain RNA > 100 ng/μL, siRNA > 10 μM.

4. It is recommended to prepare RNA pre-mixes using the dilution buffer provided with the kit to dilute the RNA stock solution.

5. The recommended concentration for RNA premix is 40 ng/μL, with a final concentration of 20 ng/μL. For co-transfection of multiple RNAs, the RNA premix concentration should be the sum of the concentrations of all RNAs.

6. Common short-chain RNAs (e.g., siRNA) typically weigh between 13 - 15 μg per 1 nmol without special modifications. The exact weight can be calculated based on the specific nucleic acid sequence and modifications.

7. The RNA/LNP ratio is crucial for LNP encapsulation. If the per-well RNA amount needs adjustment, the volume of RNA-LNP complex added to each well should be proportionally adjusted.

8. RNA-LNP complexes can be stably stored at room temperature for up to 8 h. Do not store RNA-LNP complexes under freezing conditions.

9. There is no need to replace the culture medium before or after transfection. Complete medium containing serum and antibiotics does not significantly affect transfection efficiency.

10. 6 h after transfection, standard cell culture processes, such as medium replacement, can be performed.

11. Cell conditions greatly influence transfection efficiency. It is recommended to use cells in good growth condition for transfection.

12. There are many factors affecting transfection efficiency, such as cell type, cell state and density, nucleic acid quality and concentration, ratio of transfection reagents to nucleic acids, etc. It is recommended to perform pre-experiment to determine optimal transfecti