Difference between revisions of "Part:BBa K4586020"

Latest revision as of 18:55, 11 October 2023

SV40 NLS

Part Description

Nuclear localization signals are short peptides that are responsible for the transport of nuclear protein from the cytoplasm to the nucleus to perform its function.

Usage

We used this basic part in our design to mediate the transfer of our transcription module VP64 into the nucleus to carry out its action on the target promoter that regulates the expression of our therapeutic agent (Cass12k).

Literature Characterization

The study made western blot analysis and confocal immunofluorescent analysis of 293T cells using SV40 Large T antigen.

The study used SV40 Large T Antigen (D1E9E) Rabbit mAb in the upper and β-Tubulin (D2N5G) Rabbit mAb #15115 in the lower, which were then assessed by Western blot analysis of extracts from 293 and 293T cells.

It is a confocal immunofluorescent analysis of 293T cells as they appear positive on the left and a confocal immunofluorescent analysis of HeLa cells as they appear negative on the right, and this was done using SV40 Large T antigen (D1E9E) Rabbit mAb, which appeared green, and actin filaments were labeled with DyLightTM 554 Phalloidin #13054, which appeared red.

charactrization by mathematical modeling

Presence of SV40 NIS will aid transfer of transcription factor (VP64) to the nucleus to activate (ZF21.16 minCMV) promoter to start transcription of the internal circuit that secretes the exosome's cargo to produce our engineered exosomes.

The graph shows the relation between activation of the internal domain of Syn-Notch and increasing the level of the engineered exosomes.

References

Cheng, J. et al. (2009) Semin Cancer Biol 19, 218-28.

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 10: / Line 10: @@
 ==Literature Characterization==
 The study made western blot analysis and confocal immunofluorescent analysis of 293T cells using SV40 Large T antigen.
-<html><div align="center"style="border:solid #17252A; width:50%;float:center;"><img style="                              	max-width:850px;
+<html><div align="center"style="border:solid #17252A; width:30%;float:center;"><img style="                              	max-width:850px;
 							width:75%;
 							height:auto;
@@ Line 35: / Line 35: @@
 <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%'>It is a confocal immunofluorescent analysis of 293T cells as they appear positive on the left and a confocal immunofluorescent analysis of HeLa cells as they appear negative on the right, and this was done using SV40 Large T antigen (D1E9E) Rabbit mAb, which appeared green, and actin filaments were labeled with DyLightTM 554 Phalloidin #13054, which appeared red. </span></p></div></html>
+==charactrization by mathematical modeling==
+Presence of SV40 NIS will aid transfer of transcription factor (VP64) to the nucleus to activate (ZF21.16 minCMV) promoter to start transcription of the internal circuit that secretes the exosome's cargo to produce our engineered exosomes.
+<html><div align="center"style="border:solid #17252A; width:100%;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/modeling/20.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%'>The graph shows the relation between activation of the internal domain of Syn-Notch and increasing the level of the engineered exosomes. </span></p></div></html>
 ==References==
 Cheng, J. et al. (2009) Semin Cancer Biol 19, 218-28.