Hi All,
I am planning to use 2 gRNAs and wt-CRISPR to KO my target protein from the cell lines I pick (ex. Hepatocellular carcinoma cell lines). I want to clone both gRNAs into vectors and transfect the purified plasmid. However, I am not sure what is the best way to transfect those huge vectors into target cells.
Are there any good transfection reagents for this purpose? I would like to avoid electroporation if possible.
Thank in advance for any suggestions!!
Best,
Lyra
Please go through this link.
http://www.atcc.org/~/media/Transfection%20protocols/TransfeX/HepG2%20on%20letterhead%20final.ashx
Please go through this link.
http://www.atcc.org/~/media/Transfection%20protocols/TransfeX/HepG2%20on%20letterhead%20final.ashx
Dear Lyra,
I would recommend you to try two different methods: the "classic" lipofection method that uses lipid-based transfection reagents, or the Magnetofection technology that target transfection of magnetized complexed by the way of a magnetic field.
the Magnetofection method is really efficient and recommended for primary cells and hard-to-transfect cells and was demonstrated to transfect large DNA such as BAC.
regarding CRISPR/Cas9 you can visit the following link that present all the solutions we offer for genome editing.
for any question, you can contact me via Researchgate or directly at: [email protected]
best regards,
Cedric
http://www.ozbiosciences.com/content/58-transfection-reagents-for-genome-editing
Hi Syed and Cédric,
Thank you for your information. After reading more about CRISPR/Cas9 system, I decided to directly transfect the vectors into my target cell lines by lipofectamin 3000.
I will let you know how it goes.
Best,
Lyra
Hi Lyra,
How did your transfection results turn out with Lipofectamine?
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