Transfection of Stem Cells with CRISPR Plasmids Using Lonza Kit

Centre for Cellular and Molecular Biology
Laboratory of t-RNA Transcription
Research Associate

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Company:

Lonza

Product Name:

Human Stem Cell Nucleofector Kit

Catalog Number:

VPH-5012

Our aim is to successfully transfect cancer stem cells with CRISPR plasmids to generate knock-ins of specific proteins. We transfected cancer stem cells with CRISPR plasmids and template using human stem cell nucleofector reagents. 48 h after transfection cell were sorted based on GFP expression and successfully generated single clones of knock-ins after screening.

Experimental Design and Results Summary

Application

Stem cell transfection

Starting Material

Cancer stem cells

Tips

We used nucleofector 2b device from Lonza for nucleofection

Results Summary

Successfully generated multiple clones of knock-ins with one round of transfection

DOI or PMID #

N/A

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Summary

The Good

Least toxic to cells

The Bad

N/A

The Bottom Line

Cancer stem cells are hard to transfect using lipofectamine and toxic to cells. This kit increased the efficiency of transfection and non-toxic to stem cells.

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