Part:BBa_K392001:Experience

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Applications of BBa_K392001

Through data collection, we know that ENO2 promoter has the function of driving the high-level expression of differentiated neuronal genes in vertebrates, so the study of ENO2 promoter plays a crucial role in the study of neurodegenerative diseases in vertebrates. Zebrafish,as a vertebrate, has been widely used in the study of human diseases due to its high genetic similarity with human 87 percent such as: valuable insights into disease pathogenesis,or leads to new therapeutic approaches, could be provided by high-content small molecule screens, genetic suppressor screens, observation of disease progression in real time in vivo, using fluorescent reporters to label cell populations of interest, and rapid hypothesis-testing experiments in statistically robust samples of larvae.

In the study of transgenic zebrafish, it is clearly recognized that the high expression of ENO2 can prevent neuronal degeneration by constructing neurodegeneration model, which makes an important contribution to the research on neurodegenerative diseases in the world today.The eno2 promoter will enable us to test the hypothesis that biochemical and histological changes representative of human diseases may be provoked in susceptible neuronal populations by expression of mutant transgenes.

The 12 kb eno2 promoter fragment is sufficiently small that insertion of transgene sequences into the plasmid construct is straightforward; the resulting plasmid can be grown at high copy number in Escherichia coli and DNA preparations made with ease. In addition, the relatively compact nature of the construct allows for efficient transgenesis using the I-Sce1 technique.

This figure in the literature confirm the specificity of the eno2 expression pattern demonstrated and show that the widespread hybridization pattern of the eno2 cRNA probe to brain sections is attributable to pan-neuronal expression of eno2 rather than non-specific probe hybridization.

This figure in the literature suggest that the eno2 promoter drives high-level transgene expression in adult zebrafish brain.

User Reviews

Figure 1 shows the results for the pENO2-RFP construct. Surprisingly, the RFP is localized only at the membrane, even though RFP without membrane target sequence was used.

Figure 1: Fluorescence pictures of cells expressing pENO2-RFP. The RFP is localising to the plasma membrane on the cells.

We therefore sequenced the whole construct and saw that the sequence of the promoter did not correspond to the sequence given in the registry. The sequence we found was:

TTATTATTTGTATAGTTCATCCATGCCATGTGTAATCCCAGCAGCTGTTACAAACTCAAGAAGGACCATGTGGTCTCTCTTTTCGTTGGGATCTTTCGAAAGGGCAGATTGTGTGGACAGGTAATGGTTGTCTGGTAAAnGGACAGGGCCATCGCCAATTGGAGTATTTTGTTGATAATGGTCTGCTAGTTGAACGCTTCCATCTTCAATGTTGTGTCTAATTTTGAAGTTAACTTTGATTCCATTCTTTTGTTTGTCTGCCATGATGTATACATTGTGTGAGTTATAGTTGTATTCCAATTTGTGTCCAAgAATGTTTCCATCTTCTTTAAAATCAATACCTTTTAACTCGATTCTATTAaCAAGGGTATCACCTTCAAACTTGACTTCAGCACGTGTCTTGTAGTTCC

UNIQ4b64c38af821bacf-partinfo-00000000-QINU UNIQ4b64c38af821bacf-partinfo-00000001-QINU