Question 1

What are the lipid compositions/formulation of your mRNA-LNP/siRNA-LNP?

Question 2

For standard RNA-LNPs service, what materials should I provide?

Accepted Answer

For our standard LNP formulations, you only need to provide siRNA (>= 10  nmol) / mRNA (>= 100 ug) to us.

Each synthesis will be considered one standard service, depending on the RNA preparation scale. Additional QC testing—such as UPLC for lipid composition analysis/Zeta Potential for surface charge detection of LNPs—will incur extra fees beyond the standard service charge.

Question 3

For customized RNA-LNPs service, what materials and information should I provide?

Accepted Answer

For customized LNP synthesis, customers should provide lipid materials (special lipids not included in the standard LNP formulation), formulation information (molar ratio, molecular weight of each lipid, N/P ratio or RNA/ionizable lipid W/W %), and the loading cargos: mRNA (≥100 μg/sequence)/siRNA (≥10 nmol/sequence).

Each synthesis will be considered one standard service, depending on the RNA preparation scale. Additional QC testing—such as UPLC for lipid composition analysis/Zeta Potential for surface charge detection of LNPs—will incur extra fees beyond the standard service charge.

Question 4

What are the QC tests for your RNA-LNPs?

Accepted Answer

For RNA-LNP QC tests, the specification data below apply only to the two standard formulations: SM102-LNP and MC3-LNP.

Question 5

What are recommended doses of siRNA-LNP/mRNA-LNP in vitro?

Accepted Answer

We advise customers to test various concentrations of siRNA/mRNA on their own. As a starting point, we suggest using siRNA-LNP at a final siRNA concentration of 100 nM and mRNA-LNP at 1 ng/µL of mRNA for in vitro experiment.

Question 6

What are recommended doses of siRNA-LNP/mRNA-LNP in vivo?

Accepted Answer

siRNA-LNPs: 2 mg/kg, i.v. injection, 1 injection every 3 days for 2-week period to 3 weeks period. Reporter mRNA-LNP: 0.2 mg/kg. i.v. injection, 1 injection only, 24h post i.v. imaging test.

Question 7

Any recommendations as to time points to determine alterations in the presence of the target gene?

Accepted Answer

We suggest using cell viability(>80%) to determine the optimal time points for assessing gene expression.

Question 8

What is the lead time for RNA-LNPs?

Accepted Answer

On-shelf reporter mRNA-LNP requires one working day plus shipping time, while custom RNA-LNPs typically takes 1 week, depending on the manufacturing scale.

Question 9

What is the recommended storage condition for mRNA-LNPs/siRNA-LNPs?

Accepted Answer

siRNA-LNP: 4 °C, 3 months, sterile, open inside BSC; 
mRNA-LNP: -80 °C, 3 months, avoid multiple freeze thaw cycles

Question 10

What are some alternative RNA transfection assay I can use as a control instead of LNP?

Accepted Answer

Lipofectamine™ RNAiMAX Transfection Reagent

Question 11

Do I need electroporation or other techniques to increase cell membrane permeability?

Accepted Answer

It is generally not necessary when using lipid nanoparticles. The LNPs are designed to facilitate the entry of RNA into cells without the need for additional methods like electroporation.

Question 12

Is there anyway of enriching for the cells that have taken up the RNA-LNPs?

Accepted Answer

We provide reporter mRNAs (EGFP, EYFP, and mCherry). After treatment, use fluorescence-activated cell sorting (FACS) to isolate the cells that have taken up the labeled RNA-LNP.

Question 13

For the treatment of cells, do you have a protocol on how to do that?

Accepted Answer

The RNA-LNP we provide is sterile and ready to use for both in vitro and in vivo applications. For standard intracellular transfection, simply co-culture the RNA-LNP with cells in complete medium at 37 °C and 5% CO₂ for 24 hours. For specialized applications, we do not provide a specific treatment protocol; users should optimize their own methods according to their experimental requirements.

Question 14

How to transfect cells using RNA-LNPs?

Accepted Answer

Add the RNA-LNP complex to cells and incubate for 24h at 37°C and 5% CO2. The lipid nanoparticles will fuse with the cell membrane, allowing the RNA to enter the cells.

FAQs

Frequently asked questions

What are the lipid compositions/formulation of your mRNA-LNP/siRNA-LNP?

For standard RNA-LNPs service, what materials should I provide?

For customized RNA-LNPs service, what materials and information should I provide?

What are the QC tests for your RNA-LNPs?

What are recommended doses of siRNA-LNP/mRNA-LNP in vitro?

What are recommended doses of siRNA-LNP/mRNA-LNP in vivo?

Any recommendations as to time points to determine alterations in the presence of the target gene?

What is the lead time for RNA-LNPs?

What is the recommended storage condition for mRNA-LNPs/siRNA-LNPs?

What are some alternative RNA transfection assay I can use as a control instead of LNP?

Do I need electroporation or other techniques to increase cell membrane permeability?

Is there anyway of enriching for the cells that have taken up the RNA-LNPs?

For the treatment of cells, do you have a protocol on how to do that?

How to transfect cells using RNA-LNPs?

Resources

Tool to calculate the molecular weight of your RNA/DNA samples

nmol to ug Calculator

RNA/DNA Mass to/from Moles Convertor

Primer designing tool

Recommended supplier to order custom siRNA sequences

Reccomended Negative Control siRNA

Tool to identify the DNA/RNA sequences

Tool to predict the secondary structures of DNA/RNA sequences

Recommended supplier to order custom plasmids

Tool to identify exons and introns

Tool to identify protein/peptide

Reccomended supplier to order primers

Calculator for GC content and melting temperature (Tm)

Tool to generate RNA or LNP scheme figures

Recommended supplier to order plasmids

Peptide and Protein Molecular Weight Calculator

Reporter RNA: Fluorescence emission/excitation spectrum viewer

Useful Numbers for Cell Culture

Codon optimization Tool