Difference between revisions of "Part:BBa K4586022"

Revision as of 14:45, 24 September 2023

ZF21.16-VP64

Part Description

This part is coding for our intracellular transcription module, which is formed of 3 elements: the first is zinc finger 21.16, which drives our transcription module to its promoter; the second is nuclear localization signal SV40, which mediates the entrance of the transcription unit into the nucleus; the previous two elements are linked through a G4S linker in between; and the third is VP64, which is a potent transcription factor.

Usage

This part is implemented in our design to represent the internal domain of the syn notch receptor, which controls the level of our therapeutic agent by releasing the transcription factor VP64, which triggers the expression of the internal circuit that secretes the exosomes' cargo, which is regulated by ZF 21-16 minCMV promoter activity as shown in figure 1.

Figure 1: This figure illustrates the construction of ZF21.16-VP64 that represents the internal domain of our synthetic notch receptor expressed on engineered MSC.

Literature Characterization

The study investigate the effect of activation domain (AD) and different promoters on gene expression with selection of promoter (two factor ANOVA p<0.001) and choice of AD(p<0.001)and interaction between variables(p<0.001)

So they found that mutant ZF increased gene expression with VP64 more than VP16(one-tailed Welch’s t-test,P<0.05)and with VPR more than VP64(one-tailed Welch’s t-test,P<0.01). All VPR-ZF caused the same expression in wt and mutant ZF (turkey HSD test with α = 0.05).

References

H. I., Bagheri, N., & Leonard, J. N. (2020). The COMET toolkit for composing customizable genetic programs in mammalian cells. Nature communications, 11(1), 779. https://doi.org/10.1038/s41467-019-14147-5

Sequence and Features

Assembly Compatibility:

10
COMPATIBLE WITH RFC[10]
12
COMPATIBLE WITH RFC[12]
21
COMPATIBLE WITH RFC[21]
23
COMPATIBLE WITH RFC[23]
25
COMPATIBLE WITH RFC[25]
1000
COMPATIBLE WITH RFC[1000]

@@ Line 22: / Line 22: @@
 lang=EN style='font-size:11.0pt;line-height:115%'>Figure 1: This figure illustrates the construction of ZF21.16-VP64 that represents the internal domain of our synthetic notch receptor expressed on engineered MSC. </span></p></div></html>
+==Literature Characterization==
+The study investigate the effect of activation domain (AD) and different promoters on gene expression with selection of promoter (two factor ANOVA p<0.001) and choice of AD(p<0.001)and interaction between variables(p<0.001)
+<html><div align="center"style="border:solid #17252A; width:50%;float:center;"><img style="                              	max-width:850px;
+							width:75%;
+							height:auto;
+								position: relative;
+							top: 50%;
+							left: 35%;
+							transform: translate( -50%);
+							padding-bottom:25px;
+							padding-top:25px;
+							"src="https://static.igem.wiki/teams/4586/wiki/literature-characterisation-parts/zf21-16-vp64.png">
+<p class=MsoNormal align=center style='text-align:left;border:none;width:98% ;justify-content:center;'><span
+lang=EN style='font-size:11.0pt;line-height:115%'>So they found that mutant ZF increased gene expression with VP64 more than VP16(one-tailed Welch’s t-test,P<0.05)and with VPR more than VP64(one-tailed Welch’s t-test,P<0.01). All VPR-ZF caused the same expression in wt and mutant ZF (turkey HSD test with α = 0.05).
+ </span></p></div></html>
+==References==
+H. I., Bagheri, N., & Leonard, J. N. (2020). The COMET toolkit for composing customizable genetic programs in mammalian cells. Nature communications, 11(1), 779. https://doi.org/10.1038/s41467-019-14147-5
 <!-- Add more about the biology of this part here