Generally, CRISPR-Cas9 editing leads to the formation of indels at the target site in the coding sequence of a gene. This often results in an altered reading frame (frameshift mutation) which will then lead to the formation of truncated/ non-functional protein. So, in this scenario, transcription should continue as usual and we should see a similar gene expression level as WT in the qRT-PCR. There is no doubt that transcripts will produce the truncated/ non-functional protein. Am I correct?
The outcome depends on how you design the change in the sequence of the target gene. It is possible to remove the whole gene and then it is truly gone. If it is done as you suggest, then you will get truncated/ non-functional protein.
It depends. There are "non-sense mediated decay" pathways that can remove mRNAs in the cell. But sometimes the mRNA expression levels are not changed. You'll have to figure it out empirically for your gene of interest.
- Badges
- Science topic