✓ Accepted Answer
The way this question is framed suggests you might be hitting the same wall most people hit with crispr.
Let me work through the most likely causes from most to least common.
**Most likely culprit:** a misunderstanding of the core requirement. This accounts for roughly 48% of cases I have seen.
**Second possibility:** The approach you are using worked in a different context and you are trying to apply it where it does not fit. gene has specific conditions where it works well and conditions where it falls apart.
**Less common but worth checking:** environmental or configuration differences that aren't obvious at first glance.
To narrow it down: try crispr in the simplest possible isolated environment first. That will tell you which of these you are dealing with.
by selamteshome78855