Part:BBa_K5301016
PEG10 UTR flanked-GFP
The mRNA produced by this gene can be carried by the VLP formed by PEG10. After GFP is expressed in target cells, it can be used to detect whether VLP has successfully invaded cells.
Characterization
The plasmid cargoRNA (Fig. 1) containing this gene fragment was transfected into HEK293FT cells. After 24 hours of transfection, we observed that the color of the cell medium gradually turned green, and green fluorescence was observed in the cells under fluorescence microscopy. (Fig. 2,3) These experimental results showed that PEG10 UTR flanked GFP could be expressed normally in cells, and the PEG10 UTR sequence did not affect the function of GFP.
Fig.1 Plasimd profile of cargoRNA
Fig.2 Color comparison of untransfected (left) and transfected (right) HEK293FT cell medium The medium color turned green after transfection.
Fig.3 The transfected cells emit green fluorescence (200×)
After the mRNA of PEG10 UTR flanked GFP gene fragment is transported to the target cells by PEG10 VLP, it can be translated in the target cells to express green fluorescent protein and make the cells fluoresce. (Fig. 4)
Fig.4 After being infected by PEG10 VLP exosomes, the interior of the cells fluoresced green.
Sequence and Features
- 10COMPATIBLE WITH RFC[10]
- 12COMPATIBLE WITH RFC[12]
- 21INCOMPATIBLE WITH RFC[21]Illegal BamHI site found at 263
- 23COMPATIBLE WITH RFC[23]
- 25COMPATIBLE WITH RFC[25]
- 1000INCOMPATIBLE WITH RFC[1000]Illegal BsaI site found at 307
Illegal BsaI.rc site found at 147
Illegal SapI.rc site found at 370
References
[1] Segel, M., et al., Mammalian retrovirus-like protein PEG10 packages its own mRNA and can be pseudotyped for mRNA delivery. Science, 2021. 373(6557): p. 882-889.
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