I want to completely delete or largely truncate a gene and not only achieve a functional knockout in a cell line. I can only transfect my cells with the Crispr/Cas9 sgRNA RNP and not introduce a vector construct. How can I make sure that a premature stop codon is introduced, whether I should target the promoter or just cut out the entire gene using 2 gRNAs? Any ideas?
Thanks in advance :)