Difference between revisions of "Part:BBa K4179020"
Yasmin Habib (Talk | contribs) |
Yasmin Habib (Talk | contribs) |
||
| Line 2: | Line 2: | ||
__NOTOC__ | __NOTOC__ | ||
<partinfo>BBa_K4179020 short</partinfo> | <partinfo>BBa_K4179020 short</partinfo> | ||
| + | [[Image:Hippo image.png|thumb|400px|'''Figure 1: The Hippo pathway [2]: red X showing decursin's effect ''']] | ||
The composite part comprises a Blasticidin S deaminase [https://parts.igem.org/Part:BBa_K1943015 (BBa_K1943015)], which is a Blasticidin resistance gene with the Kozak sequence, under the CMV constitutive promoter [https://parts.igem.org/Part:BBa_I712004 (BBa_I712004)]. Downstream to it an SV40PolyA terminator [https://parts.igem.org/Part:BBa_K4235020 (BBa_K4235020)] is found. Together in this composite, a cTNT promoter [https://parts.igem.org/Part:BBa_K4179021 (BBa_K4179021)] is cloned downstream to the 8XGTIIC motifs [https://parts.igem.org/Part:BBa_K4179017 (BBa_K4179017)], followed by the luciferase gene [https://parts.igem.org/Part:BBa_I712019 (BBa_I712019)] and finally the SV40 Poly(A) [https://parts.igem.org/Part:BBa_K4235020 (BBa_K4235020)]. | The composite part comprises a Blasticidin S deaminase [https://parts.igem.org/Part:BBa_K1943015 (BBa_K1943015)], which is a Blasticidin resistance gene with the Kozak sequence, under the CMV constitutive promoter [https://parts.igem.org/Part:BBa_I712004 (BBa_I712004)]. Downstream to it an SV40PolyA terminator [https://parts.igem.org/Part:BBa_K4235020 (BBa_K4235020)] is found. Together in this composite, a cTNT promoter [https://parts.igem.org/Part:BBa_K4179021 (BBa_K4179021)] is cloned downstream to the 8XGTIIC motifs [https://parts.igem.org/Part:BBa_K4179017 (BBa_K4179017)], followed by the luciferase gene [https://parts.igem.org/Part:BBa_I712019 (BBa_I712019)] and finally the SV40 Poly(A) [https://parts.igem.org/Part:BBa_K4235020 (BBa_K4235020)]. | ||
| Line 7: | Line 8: | ||
===Usage and Biology=== | ===Usage and Biology=== | ||
| − | The team of Technion 2022 used this composite as a part of their cell line measurement system called OraCell. This biological system quantifies their manufactured metabolite-decursin by utilizing its effect on the Hippo pathway which is a signaling pathway that regulated proliferation and apoptosis in mammalian cells [1]. TEAD transcriptional factors bind to the 8XGTIIC motifs and regulate the expression of the luciferase gene which is cloned downstream to these motifs. Luciferase is used as a reporter gene that emits luminescence. Decursin inhibits expression of luciferase gene by inhibiting the Hippo pathway. Thus, the quantity of this metabolite is measured according to the luminescence values, by using a calibration curve. This composite is transfected to CHO (Chinese Hamster Ovarian) cells by stable integration. The role of the Blasticidin S deaminase here is to accomplish a successful selection for positive cells after transfection. To read more about the OraCell measurement tool, visit the team's wiki page. | + | The team of Technion 2022 used this composite as a part of their cell line measurement system called OraCell. This biological system quantifies their manufactured metabolite-decursin by utilizing its effect on the Hippo pathway which is a signaling pathway that regulated proliferation and apoptosis in mammalian cells [1]. TEAD transcriptional factors bind to the 8XGTIIC motifs and regulate the expression of the luciferase gene which is cloned downstream to these motifs. Luciferase is used as a reporter gene that emits luminescence. Decursin inhibits expression of luciferase gene by inhibiting the Hippo pathway (figure 1). Thus, the quantity of this metabolite is measured according to the luminescence values, by using a calibration curve. This composite is transfected to CHO (Chinese Hamster Ovarian) cells by stable integration. The role of the Blasticidin S deaminase here is to accomplish a successful selection for positive cells after transfection. To read more about the OraCell measurement tool, visit the team's wiki page. |
| + | |||
| + | |||
| + | ===Characterization=== | ||
| + | |||
| + | |||
| + | |||
| − | |||
| − | |||
<!-- --> | <!-- --> | ||
<span class='h3bb'> | <span class='h3bb'> | ||
| − | Sequence and Features | + | ===Sequence and Features=== |
<partinfo>BBa_K4179020 SequenceAndFeatures</partinfo> | <partinfo>BBa_K4179020 SequenceAndFeatures</partinfo> | ||
| + | |||
| + | ==References== | ||
| + | 1.Yu, F. X., & Guan, K. L. (2013). The Hippo pathway: Regulators and regulations. In Geneand Development (Vol. 27, Issue 4, pp. 355–371). doi: 10.1101/gad.210773.112 | ||
| + | 2. Liu, Q., Liu, X., & Song, G. (2021). The Hippo Pathway: A Master Regulatory Network Important in Cancer. Cells, 10(6), 1416. https://doi.org/10.3390/cells10061416 | ||
<partinfo>BBa_K4179020 parameters</partinfo> | <partinfo>BBa_K4179020 parameters</partinfo> | ||
<!-- --> | <!-- --> | ||
Revision as of 15:00, 8 October 2022
Luc_Blast_OraCell
The composite part comprises a Blasticidin S deaminase (BBa_K1943015), which is a Blasticidin resistance gene with the Kozak sequence, under the CMV constitutive promoter (BBa_I712004). Downstream to it an SV40PolyA terminator (BBa_K4235020) is found. Together in this composite, a cTNT promoter (BBa_K4179021) is cloned downstream to the 8XGTIIC motifs (BBa_K4179017), followed by the luciferase gene (BBa_I712019) and finally the SV40 Poly(A) (BBa_K4235020).
Usage and Biology
The team of Technion 2022 used this composite as a part of their cell line measurement system called OraCell. This biological system quantifies their manufactured metabolite-decursin by utilizing its effect on the Hippo pathway which is a signaling pathway that regulated proliferation and apoptosis in mammalian cells [1]. TEAD transcriptional factors bind to the 8XGTIIC motifs and regulate the expression of the luciferase gene which is cloned downstream to these motifs. Luciferase is used as a reporter gene that emits luminescence. Decursin inhibits expression of luciferase gene by inhibiting the Hippo pathway (figure 1). Thus, the quantity of this metabolite is measured according to the luminescence values, by using a calibration curve. This composite is transfected to CHO (Chinese Hamster Ovarian) cells by stable integration. The role of the Blasticidin S deaminase here is to accomplish a successful selection for positive cells after transfection. To read more about the OraCell measurement tool, visit the team's wiki page.
Characterization
Sequence and Features
- 10COMPATIBLE WITH RFC[10]
- 12COMPATIBLE WITH RFC[12]
- 21COMPATIBLE WITH RFC[21]
- 23COMPATIBLE WITH RFC[23]
- 25INCOMPATIBLE WITH RFC[25]Illegal NgoMIV site found at 213
- 1000INCOMPATIBLE WITH RFC[1000]Illegal SapI.rc site found at 1090
References
1.Yu, F. X., & Guan, K. L. (2013). The Hippo pathway: Regulators and regulations. In Geneand Development (Vol. 27, Issue 4, pp. 355–371). doi: 10.1101/gad.210773.112 2. Liu, Q., Liu, X., & Song, G. (2021). The Hippo Pathway: A Master Regulatory Network Important in Cancer. Cells, 10(6), 1416. https://doi.org/10.3390/cells10061416