I have transfected neurons using electroporation before playing plasmid with Cas9 which makes a single double stranded cut. But I am not able to see any results. Is there a way I can delete the whole gene using CRISPR in primary neuron culture?
Primary neurons are very difficult to transfect, and electroporation likely damages the neurons; you can try to package your Cas9 into a lentiviral vector; another approach, if you purchase the knockout mouse with your target gene (if it exists), you can isolate primary neurons from its brain with knockout already present.
Thanks for the advice. The knock out mouse is not available but I will try the lentivirus .
Thanks for the advice. The knock out mouse is not available but I will try the lentivirus .
Tripti
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