Hi, I produced a stable expressing Cas9 cell line. After transducing the cells, I checked the protein expression of my protein of interest and I understood that the transduced cells expressed a protein of interest less that the non-transduced cells. Does anybody have the same experience?
I have seen this in cells growing in puromycin, and I believe it's related to the puro. It should be fine as long as your experiments are conducted using a control cell line that's growing in the same selection antibiotic(s) and has the same plasmid with a non-targeting insert.
It is possible that the expression of Cas9 in your stable expressing cell line could affect the expression of other proteins in the transduced cells. Cas9 is an enzyme that plays a central role in the CRISPR/Cas9 genome editing system. It recognizes and cuts specific sequences of DNA, and this process can potentially affect the expression of other proteins in the cell.
There are several potential mechanisms by which the expression of Cas9 could affect the expression of other proteins in transduced cells. For example, Cas9 might bind to and cleave the regulatory sequences of certain genes, disrupting the normal regulation of gene expression. Alternatively, Cas9 might cause DNA damage or DNA double-strand breaks that could affect the expression of other proteins in the cell.
It is also worth noting that the expression of Cas9 might be influenced by other factors in the cell, such as the presence of specific signaling pathways or transcription factors. Therefore, it is possible that the observed differences in protein expression between transduced and non-transduced cells could be due to the combined effects of Cas9 expression and other factors in the cell.
To determine the specific mechanism by which Cas9 expression is affecting the expression of other proteins in your stable expressing cell line, it may be necessary to conduct further experiments. This could include analyzing the expression of specific genes or proteins using techniques such as RT-qPCR or western blotting, or examining the effects of specific signaling pathways or transcription factors on Cas9 expression and protein expression.
- Badges
- Science topic
- Similar topics
- Biological Science
- Microbiology
- Virology
- Virus