Pre-Synthesized mRNA for Cell Manipulation

January 05, 2016

Brigham and Women's Hospital
Pathology
Postdoctoral Fellow

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Company:

Trilink

Product Name:

Enhanced Green Fluorescent Protein mRNA

Catalog Number:

L-6301

Preparing good quality mRNA for transfection is difficult due to specific requirements of the polyA tail and mRNA instability. I tested the pre-synthesized mRNA from Trilink and it worked very well when transfected into T cells. The expression was high at 24 hours and stayed for 5 days after transfection.

Experimental Design and Results Summary

Application

Transfection of T cells

Starting Material

Purified human T cells

Protocol Overview

About 5e6 T cells were electroporated with 2ug of GFP mRNA using the Neon electroporator. The cells were then rested in 10%RPMI supplemented with 10ng/ml IL2 for the next 4-5 days.

Tips

Keep cells healthy by feeding IL2 and splitting them when confluent.

Results Summary

The GFP expression was very bright at 24 hours post electroporation.

Additional Notes

None

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Summary

The Good

Good quality mRNA for transfections

The Bad

Expensive

The Bottom Line

The mRNA was stable for a long time and worked very well with transfecting T cells which are very difficult to transfect with plasmids. A god way to engineer cells without using lentiviruses.

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