What are the Challenges in Knockout and Knockdown Experiments?

Despite their utility, these experiments come with challenges:

Off-target effects: Both CRISPR-Cas9 and RNAi can sometimes affect genes other than the intended target, leading to misleading results.
Incomplete Knockdown: RNAi may not completely reduce gene expression, making it difficult to interpret the results.
Compensatory Mechanisms: Cells can activate alternative pathways to compensate for the loss of a gene, complicating the analysis.
Ethical Considerations: Especially in vertebrate models, ethical concerns must be addressed when creating knockout or knockdown organisms.