Difference between revisions of "Part:BBa K5036020"

Latest revision as of 08:42, 1 October 2024

Yes Associated Protein-1 (YAP)

Part Description

It is a transcriptional coactivator that plays a crucial role in various biological processes, including organ development, tissue regeneration, and cancer progression. It is a key component of the Hippo pathway which is a signaling pathway that regulates cell growth, proliferation, and differentiation. Furthermore, YAP-1 has a function in controlling stem cell activity. Ensuring that there is a generation of new cells and their specialization to maintain a steady cell supply.

Usage

YAP-1 arrives to the nucleus where YAP-1 stimulates transcription factors, modifying the expression of genes implicated in cell growth so it increases cell self-renewal which increase the rate of wound healing minimizing the extent of the scar. in our model We increased YAP-1 levels in our engineered MSCs and released YAP-1-containing exosomes to promote wound healing. We used a dCas9(C/N)-TF-VEGFR1/2 receptor and a TID switch to control YAP-1 production.

this figure illustrates the structure of yes associated protein-1 (YAP-1) .

Characterization by Mathematical Modeling

The model provides the kinetics of YAP-1 either from increasing its transcription level using different transcriptional activators or initiating its translation from our conditioned translation initiation device (TID) switch. It is all based on parametric values from literature

Graph (1). Illustrates the relation between activation levels of different transcriptional activators (CMV trans-enhancer , VP64 and GAL4) for Increasing transcription level for YAP-1 .

Graph(1). Illustrates the relation between circulation form activity of our switch (Yellow line) and their ability for YAP-1 production (Black line) upon their binding to MMP-9 .

Literature characterization

This study examined the importance of a molecule called STAT3 in gene activation controlled by YAP-1 in liver cancer cells (HuH7 hepatoma cells). We achieved this by STAT3 pharmacological inhibition and by genetic silencing. they tested these effects under normal conditions and when YAP-1 levels were artificially increased

Figure A shows that while treating liver cancer cells (HuH7) with a STAT3 inhibitor (Stattic) slightly reduced Snail gene activity, it caused a significant decrease in the production of CTGF and Cyr61 genes (known YAP-1/TEAD targets). This effect was also observed when STAT3 production was lowered using a different method (siRNA knockdown) – as shown in Figure C. Interestingly, both experiments also showed a decrease in YAP-1 itself (RNA and protein levels) – Figures B and D. This suggests STAT3 might regulate YAP-1 gene expression. However, Static treatment still reduced the activity of 64 YAP-1 target genes even when YAP-1 levels were artificially increased (Figure E and F). This implies STAT3 might control YAP-1's ability to activate genes, suggesting a complex regulatory role by STAT3 on YAP-1 activity.

Reference

Consalvi, V. (2024). Characterization of the transcriptional complexes enrolled by Yes-associated protein (YAP) in hepatoma cells and their regulation by ERK5/MAPK.

Xu X, Gao J, Dai W, Wang D, Wu J, Wang J. Gene activation by a CRISPR-assisted trans enhancer. Elife. 2019 Apr 11;8:e45973. doi: 10.7554/eLife.45973. PMID: 30973327; PMCID: PMC6478495.

Morita S, Horii T, Kimura M, Hatada I. Synergistic Upregulation of Target Genes by TET1 and VP64 in the dCas9-SunTag Platform. Int J Mol Sci. 2020 Feb 25;21(5):1574. doi: 10.3390/ijms21051574. PMID: 32106616; PMCID: PMC7084704.

Shao, J., Li, S., Qiu, X. et al. Engineered poly(A)-surrogates for translational regulation and therapeutic biocomputation in mammalian cells. Cell Res 34, 31–46 (2024). https://doi.org/10.1038/s41422-023-00896-y ‏

Sequence and Features