Difference between revisions of "Part:BBa K1699001"

Revision as of 13:44, 10 September 2015

Human short TERT promoter

Human short TERT promoter.

hTERT (human Telomerase Reverse Transcriptase) is a human promoter which controls the transcription of Telomerase, a gene highly expressed in cancer cells, and not in healthy ones (1).

Usage and Biology

hTERT promotes the transcription of the catalytic subunit of telomerase. Telomerase elongates the ends of chromosomes, regions called telomers. hTERT is not active in somatic cells and is highly active in most if not all cancer cell types. Therefore, the expression of an exogenous gene under the control of hTERT promoter will likely occur in cancer cells only. We have used this promoter as part of a two promoter system (the other one being survivin promoter, also highly active in cancer cells), in order drive the expression, and subsequently activate the core of our system (based on CRISPR?Cas9 technology) exclusively in cancer cells.

Characterization

This part was used by BGU 2015 team in a couple of constructs. Promoter activation worked in all of them.
1. hTERT GFP - robust transcription of GFP in all HepG2 cells. None detected in control (human fibroblasts).
2. hTERT dCas9-VP64 - activation system including gRNA under survivin promoter. Activation system worked in cancer cells and not in healthy ones.

References

1. The telomerase reverse transcriptase promoter drives efficacious tumor suicide gene therapy while preventing hepatotoxicity encountered with constitutive promoters http://www.nature.com/gt/journal/v8/n7/pdf/3301421a.pdf

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: @@
 ===Usage and Biology===
-hTERT promotes the transcription of the catalytic subunit of telomerase. Telomerase elognates the ends of chromosomes, regions called telomers. hTERT is not active in somatic cells and appears to be highly active in most if not all cancer cells, therefor, cells that will express an exogenic gene under hTERT are likely to be cancerous.
+hTERT promotes the transcription of the catalytic subunit of telomerase. Telomerase elongates the ends of chromosomes, regions called telomers. hTERT is not active in somatic cells and is highly active in most if not all cancer cell types. Therefore, the expression of an exogenous gene under the control of hTERT promoter will likely occur in cancer cells only.
-We have used this promoter as part of a two promoter system, in order to detect whether a cell was a cancer cell.
+We have used this promoter as part of a two promoter system (the other one being survivin promoter, also highly active in cancer cells), in order drive the expression, and subsequently activate the core of our system (based on CRISPR?Cas9 technology) exclusively in cancer cells.
 ===Characterization===