What are recommended doses of siRNA-LNP/mRNA-LNP in vitro?
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.
What are recommended doses of siRNA-LNP/mRNA-LNP in vivo?
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.
Do you provide service of custom oligos synthesis?
We recommend that customers purchase oligos, and we can assist with the encapsulation process.
How to transfect cells using RNA-LNPs?
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
Any recommendations as to time points to determine alterations in the presence of the target gene?
We suggest using cell viability(>80%) to determine the optimal time points for assessing gene expression.
Do I need electroporation or other techniques to increase cell membrane permeability?
How do I detect EGFP/EYFP/mCherry mRNA after transfection?
Do you have any siRNA available to give as a control?
What are some alternative RNA transfection kits I can use as a control instead of LNP?
For RNA-LNPs service, what materials should I provide?
You only need to provide siRNA (>= 3 nmol) / mRNA (>= 50 ug) to us.
For custom RNA synthesis, what materials I should provide?
What are the differences between your RNA-LNP kits and your formal RNA-LNP service?
For the RNA-LNP Kit, the LNP is made using manual pipetting, which may result in variations in LNP quality. For our formal RNA-LNP service, we use a microfluidic device and flow control system to prepare the LNP, followed by post-purification and QC tests using various instruments.
Any tricks that may not be in the RNA-LNP Kit protocol?
Is there anyway of enriching for the cells that have taken up the RNA-LNPs?
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.
For the treatment of cells, do you have a protocol on how to do that?
We don’t have a specific protocol for the treatment of cells. Users should develop their own method tailored to their experimental needs.
Resources
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