I been wondering if genes when stable transfected become regulated by the cell, or if the gene is constitutively express and translated and the protein previously made is degraded at a slower rate which allows protein accumulation. I have been wondering these since there need to reach an equilibrium in either case for the transfected gene (and the subsequent protein) and not to become toxic to the cell. I hope someone could direct me to a paper that explains this.
I ask this question because if a protein is inhibited by post-translational modification and is perpetually recycle (meaning the older protein is being ubiquitinated as new transcript are giving rise to new proteins) this means that the proteins are technically always active. However, if the transfected stable gene is regulated somehow, means that protein reaches a limit and all in time will become inhibited by this hypothetical post-translational modification. Then if the protein in question is naturally being regulated in transcription factors specifically present in certain cell lines (myeloid for example), but not in others. This could mean that the protein would not have activity in non-myeloid cells even if it's express since there is no way to activated unless other proteins that could undo the previous modification are also supplied.
I am thinking correctly, or this is just crazy talk?
Thank you, and stay safe
Hello, Carlos!
For exzmple,
PU.1 is a hematopoietic master transcription factor, predominantly expressed in immature myeloid cells and B cells, and downregulation of this factor is important in various hematological malignancies (46,47). To identify downstream target genes of PU.1, the authors generated cell lines expressing reduced levels of PU.1 by stable transfection of PU.1 short inhibitory RNAs into K562 human myeloid leukemia cells (K562PU.1KD cells) and PU.1-overexpressing K562 cells (K562PU.1OE cells). Dual microarray analyses were performed using these cell lines. Notably, the expression levels of all the functional MT isoforms expressed in humans (MT-1A, -B, -E, -F, -G, -H and -X and MT-2) were increased by varying degrees in the K562PU.1KD cells.
Article Positive and negative regulators of the metallothionein gene (Review)
Stable transfection can integrate the transfected DNA into the cell genome and transform it into the offspring that is passed on to the cell, thereby realizing the long-term introduction of foreign DNA into the cell and achieving continuous expression. Whether there is a change in degradation needs to consider whether the gene is related to the protein degradation pathway
learn more about transfection, please visit https://transfection.bocsci.com/
- Badges
- Science topic
- Similar topics
- Biological Science
- Molecular Cell Biology
- Culture Cells
- Cell Line