BBa_K1722001 1 shTERT shTERT is a cancer cell specific promoter with high efficiency. 2015-08-27T11:00:00Z 2015-09-01T06:24:21Z The telomerase reverse transcriptase promoter can be found in human cancer cells. In our experiment, we got the part from Shenzhen second people's hospital. Additionally, we did our system's function verification in Shenzhen second people's hospital. Telomerase reverse transcriptase??(abbreviated to??TERT, or??hTERT??in humans) is a catalytic subunit of the??enzyme??telomerase, which, together with the??telomerase RNA component??(TERC), comprises the most important unit of the telomerase complex. The telomerase is a ribonucleoprotein enzyme to which multiple functions have been attributed, the most important of these is the maintenance of the telomere which is related with cellular immortalization and cancer. 85% of human tumors have telomerase activity, that in normal cells goes undetected. These characteristics make the telomerase an attractive target for chemotherapy. TERT??promoter??mutations were highly frequent in glioblastoma (83.9%), urothelial carcinoma (64.5%), oligodendroglioma (70.0%), medulloblastoma (33.3%) and hepatocellular carcinoma (31.4%). C228T and C250T were the most common mutations. In urothelial carcinoma, several novel rare mutations were identified. TERT??promoter??mutations were absent in gastrointestinal stromal tumour (GIST), thymic epithelial tumours, gastrointestinal leiomyoma, gastric schwannoma, cholangiocarcinoma, gastric and pancreatic cancer. TERT??promoter??mutations highly correlated with upregulated TERT mRNA expression and??telomerase??activity in adult gliomas. These mutations differentially enhanced the transcriptional activity of the TERT core??promoter.TERT??promoter??mutations are frequent in multiple tumour types and have similar distributions in Chinese cancer patients. The functional significance of these mutations reflect the importance to telomere maintenance and hence tumourigenesis, making them potential therapeutic targets. More??importantly , we mutate the normal TERTp into a new TERTp with high-efficency of the promotion so as to make our system work??efficiently. In the experiment, we change 4 base of the TERTp gene. Eventually, The telomerase reverse transcriptase promoter can specifically promotes with the identification of telomerase reverse transcriptase, which means it can only promote in cancer cells.In our system, we use TERTp to promote only in cancer cells. Together with bladder-specific hUPII promoter we can achieve the precision of system???s work in bladder cancer cells.??In our experiment, we constructed three and two plasmids before and after two times to verify the function using high-efficency TERTp.The TERTp can be used to promote in cancer cells. Similar to our system, alike synthesizing gene circuits are also expected to be one of the promising approaches to the treatment to other cancer. false false _2142_ 26634 26635 9 false We designed the following primers and amplified hTERT promoter from the vector psi-Check2: Sequence(up)CCGGAATTCGGCACCTCCCTCGGGTTAG Sequence(down)TGCACTGCAGACTAGTCGCGTGGGTGGCCG. By incorporating these primers into hTERT promoter, the promoter is flanked by the iGEM prefix and suffix after amplification. false Fang Shu BBa_K1722004 1 aa-tRNA This is a kind of aminoacyl-tRNA(aa-tRNA). 2015-08-25T11:00:00Z 2015-09-07T08:29:17Z It came from barchaebacteria. This is a kind of aminoacyl-tRNA, which has a diffeent structure compared with classic tRNA.The tRNA can only carry a specific amino acid, and is non natural amino acids.It has special anticodons(CUA),which can identify the termination codon(UAG), and then adds the unnatural amino acid carried own into the polypeptide chain to.So that the position of the termination codon corresponds to the insertion of an unnatural amino acid, which allows the protein to continue to be translated. The aminoacyl-tRNA came from barchaebacteria,and it is generally applied to unnatural amino acid orthogonal system. In our project, the aminoacyl-tRNA was selected by directed evolution. false false _2142_ 20403 20403 9 false Because itonly has 72 bases, and the traditional PCR method is not applicable for this,we get his by gene synthesising. false Yongyi Wang BBa_K1722011 1 shTERTtRNA shTERT+tRNA(CUA) Composite 2015-09-01T11:00:00Z 2015-09-07T09:03:03Z Both shTERT promoter and tRNA are achieved from Shenzhen Second People's Hospital. hTERT, which is short for human telomerase reverse transcriptase, is a cancer-cell specific promoter. It can be activated inside cancer cells with no effect on normal cells. By mutating four base pairs of hTERT sequence, we achieved an improved promoter with higher promote efficiency named super hTERT(shTERT). As an important component of human telomerase, hTERT express only in tumor cells and other immortal cells which are telomerase positive. Only in tumor cells that can express TERT can the promoter being activated to realize targeted expression of effector gene. We constructed another gene downstream of shTERT called tRNA, which encodes an unusual transfer RNA(tRNA) with a CUA anticodon. The tRNA(CUA) that is produced can be charged with Acetyllysine(Ack) with the help of Acetyllysine tRNA synthetase(AckRS). It is also the first step in translating UAG amber codon. In our project, when shTERT is activated inside bladder cancer cell, downstream tRNA sequence can be expressed to produce tRNA with CUA as its anticodon. Together with AckRS, tRNA can carry Ack and pair with the amber UAG stop codon of the mRNA chain to completely produce the interest protein. false false _2142_ 20403 26634 9 false Both shTERT promoter and tRNA are achieved from Shenzhen Second People's Hospital. We designed primers and amplified the gene sequences from psi-Check2 vector. Using 3A Assembly method, we constructed hTERT and tRNA in pSB1C3. false Zhiwei Zhang component2442689 1 BBa_K1722001 component2442690 1 BBa_K1722004 annotation2442689 1 BBa_K1722001 range2442689 1 1 454 annotation2442690 1 BBa_K1722004 range2442690 1 463 534 BBa_K1722011_sequence 1 ggcccctccctcgggttaccccacagcctaggccgattcgacctctctccgctggggccctcgctggcgtccctgcaccctgggagcgcgagcggcgcgcgggcggggaagcgcggcccagacccccgggtccgcccggagcagctgcgctgtcggggccaggccgggctcccagtggattcgcgggcacagacgcccaggaccgcgctccccacgtggcggagggactggggacccgggcacccgtcctgccccttcaccttccagctccgcctcctccgcgcggaccccgccccgtcccgaccccttccgggtttccggcccagccccttccgggccctcccagcccctccccttcctttccgcggccccgccctctcctcgcggcgcgagtttcaggcagcgctgcgtcctgctgcgcacgtgggaagccctggccccggccacccccgcgtactagagggaaacctgatcatgtagatcgaatggactctaaatccgttcagccgggttagattcccggggtttccgcca BBa_K1722004_sequence 1 ggaaacctgatcatgtagatcgaatggactctaaatccgttcagccgggttagattcccggggtttccgcca BBa_K1722001_sequence 1 ggcccctccctcgggttaccccacagcctaggccgattcgacctctctccgctggggccctcgctggcgtccctgcaccctgggagcgcgagcggcgcgcgggcggggaagcgcggcccagacccccgggtccgcccggagcagctgcgctgtcggggccaggccgggctcccagtggattcgcgggcacagacgcccaggaccgcgctccccacgtggcggagggactggggacccgggcacccgtcctgccccttcaccttccagctccgcctcctccgcgcggaccccgccccgtcccgaccccttccgggtttccggcccagccccttccgggccctcccagcccctccccttcctttccgcggccccgccctctcctcgcggcgcgagtttcaggcagcgctgcgtcctgctgcgcacgtgggaagccctggccccggccacccccgcg igem2sbol 1 iGEM to SBOL conversion Conversion of the iGEM parts registry to SBOL2.1 Chris J. Myers James Alastair McLaughlin 2017-03-06T15:00:00.000Z