Hi everyone,
Thanks for your time. I am planning to knockout a gene by targeted insertion of RFP into the exon 1 by CRISPR cas mediated HDR. Since insertion of the promoter less RFP coding sequences gets integrated into the middle of the EXON 1 of the gene. This may lead to production of fusion protein. One side, i am pretty sure that the expression of the gene which i want to knock down will be successful. But, will the RFP expressed as fusion protein with the part of the EXON 1 in their upstream using the endogenous promoter.
Please enlighten me!!!