Difference between revisions of "Part:BBa K2989000"
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| − | + | HULC is a liver cancer-specific lncRNA that is Highly Up-regulated in Liver Cancer (HULC) and is one of the most up-regulated genes in HCC. In 76% of hepatocellular carcinoma tissues, the expression level of HULC was up-regulated by 33 times [8] compared with normal liver tissue. This enables HULC to be a HCC specific promoter. | |
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| + | ===Usage=== | ||
| + | We used this Hulc promoter as the upstream control element of our "open" plasmid, and with the promoter-specific ability of the liver cancer cells, the normal cells are more likely to not express the following CeRNA. The expression of the CeRNA is only expressed in the liver cancer tumor cells, and the competitive binding of the miRNA promotes the expression of the HBsAg fragment, thereby eliciting an antitumor response. | ||
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| + | https://2019.igem.org/wiki/images/0/01/T--Nanjing_NFLS--Parts_BBa_K2989000.jpg | ||
| + | Table1 Relative promoter activity of HULC and SV40 promoters<br/> | ||
| + | Vector HepG2 293T<br/> | ||
| + | PGL3-Basic 22.02 1.32<br/> | ||
| + | PGL3-hulc 237.38 130.39<br/> | ||
| + | PGL3-Control 2782.88 3318.18<br/> | ||
| + | The data is the mean of the relative Luc activity RL/RL (%) measured.<br/> | ||
| + | The PGL3-Basic is the negative control group while the PGL3-Control is the positive control group. From the table shown above, the PGL3-Basic group without any promoter showed a low relative Luc activity, indicating that the entire detection system exhibited no interference. The PGL3-Control group with SV40 promoter has good priming performance in both engineered HEK293T cell line and tumor cell HepG2 cells: maintains a good value of 3000% of relative Luc activity. However, it does not have a tumor-specific selection ability. We found that the Hulc promoter's ability to initiate tumor cells is twice that in normal cells, indicating that Hulc is an ideal promoter in the AND GATE system. We proved that Hulc has tumor-specific activity and therefore, we chose Hulc in a specific regulation system. | ||
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| + | ===Biology=== | ||
| + | Highly upregulated in liver cancer (HULC) is a highly expressed lncRNA found in liver cancer tissues. Related studies have shown that HULC is highly expressed in hepatitis B virus (HBV)-associated cirrhosis, which can directly down-regulate the tumor suppressor gene P18, increase the number of regulatory T lymphocytes, and increase the transforming growth factor (transforming growth factor). Factor, TGF) β level, promotes the process of immune response to liver fibrosis. On the other hand, the expression of hulc received transcript levels and down-regulated its activity in interaction with miR-372; the low expression of miR-372 repressed its inhibition of the target protein kinase A catalytic subunit β, induction Phosphorylation of cyclic adenosine CREB; phosphorylated CREB can specifically bind to the CREB binding site in the promoter of HULC gene, maintain the open state of the chromosome structure in the promoter of HULC gene, and increase the transcription of HULC gene and form "Positive feedback adjustment". | ||
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Latest revision as of 02:28, 22 October 2019
hulc promoter
HULC is a liver cancer-specific lncRNA that is Highly Up-regulated in Liver Cancer (HULC) and is one of the most up-regulated genes in HCC. In 76% of hepatocellular carcinoma tissues, the expression level of HULC was up-regulated by 33 times [8] compared with normal liver tissue. This enables HULC to be a HCC specific promoter.
Usage
We used this Hulc promoter as the upstream control element of our "open" plasmid, and with the promoter-specific ability of the liver cancer cells, the normal cells are more likely to not express the following CeRNA. The expression of the CeRNA is only expressed in the liver cancer tumor cells, and the competitive binding of the miRNA promotes the expression of the HBsAg fragment, thereby eliciting an antitumor response.
Table1 Relative promoter activity of HULC and SV40 promoters
Vector HepG2 293T
PGL3-Basic 22.02 1.32
PGL3-hulc 237.38 130.39
PGL3-Control 2782.88 3318.18
The data is the mean of the relative Luc activity RL/RL (%) measured.
The PGL3-Basic is the negative control group while the PGL3-Control is the positive control group. From the table shown above, the PGL3-Basic group without any promoter showed a low relative Luc activity, indicating that the entire detection system exhibited no interference. The PGL3-Control group with SV40 promoter has good priming performance in both engineered HEK293T cell line and tumor cell HepG2 cells: maintains a good value of 3000% of relative Luc activity. However, it does not have a tumor-specific selection ability. We found that the Hulc promoter's ability to initiate tumor cells is twice that in normal cells, indicating that Hulc is an ideal promoter in the AND GATE system. We proved that Hulc has tumor-specific activity and therefore, we chose Hulc in a specific regulation system.
Biology
Highly upregulated in liver cancer (HULC) is a highly expressed lncRNA found in liver cancer tissues. Related studies have shown that HULC is highly expressed in hepatitis B virus (HBV)-associated cirrhosis, which can directly down-regulate the tumor suppressor gene P18, increase the number of regulatory T lymphocytes, and increase the transforming growth factor (transforming growth factor). Factor, TGF) β level, promotes the process of immune response to liver fibrosis. On the other hand, the expression of hulc received transcript levels and down-regulated its activity in interaction with miR-372; the low expression of miR-372 repressed its inhibition of the target protein kinase A catalytic subunit β, induction Phosphorylation of cyclic adenosine CREB; phosphorylated CREB can specifically bind to the CREB binding site in the promoter of HULC gene, maintain the open state of the chromosome structure in the promoter of HULC gene, and increase the transcription of HULC gene and form "Positive feedback adjustment".
Sequence and Features
- 10COMPATIBLE WITH RFC[10]
- 12COMPATIBLE WITH RFC[12]
- 21COMPATIBLE WITH RFC[21]
- 23COMPATIBLE WITH RFC[23]
- 25COMPATIBLE WITH RFC[25]
- 1000COMPATIBLE WITH RFC[1000]