@article{604bbe54f7b3431bb1bb375f2af0bb19,
title = "Redesigned reporter gene for improved proton exchange-based molecular MRI contrast",
abstract = "Reporter gene imaging allows for non-invasive monitoring of molecular processes in living cells, providing insights on the mechanisms underlying pathology and therapy. A lysine-rich protein (LRP) chemical exchange saturation transfer (CEST) MRI reporter gene has previously been developed and used to image tumor cells, cardiac viral gene transfer, and oncolytic virotherapy. However, the highly repetitive nature of the LRP reporter gene sequence leads to DNA recombination events and the expression of a range of truncated LRP protein fragments, thereby greatly limiting the CEST sensitivity. Here we report the use of a redesigned LRP reporter (rdLRP), aimed to provide excellent stability and CEST sensitivity. The rdLRP contains no DNA repeats or GC rich regions and 30% less positively charged amino-acids. RT-PCR of cell lysates transfected with rdLRP demonstrated a stable reporter gene with a single distinct band corresponding to full-length DNA. A distinct increase in CEST-MRI contrast was obtained in cell lysates of rdLRP transfected cells and in in vivo LRP expressing mouse brain tumors (p= 0.0275 , n = 10).",
author = "Or Perlman and Hirotaka Ito and Gilad, {Assaf A.} and McMahon, {Michael T.} and Chiocca, {E. Antonio} and Hiroshi Nakashima and Farrar, {Christian T.}",
note = "Funding Information: The work was supported by the US National Institutes of Health Grants R01-CA203873, P41-RR14075, as well as by 1R01NS110942-01A1 and P01 CA163205 (EAC and HN) and NIH R01 NS098231; R01 NS104306 and P41 EB024495 (AAG). The Brigham and Women{\textquoteright}s Small Animal Imaging Laboratory (SAIL) was funded by a G20 Grant (1G20RR031051-01) as part of the American Recovery and Reinvestment Act as a part of the construction of the Brigham and Women{\textquoteright}s MRI Research Center (BWMRC). The 7T Bruker Small Bore Animal Magnet was partially funded by an S10 Grant (1S10OD010705-01) through the National Institutes of Health. This project has received funding from the European Union{\textquoteright}s Horizon 2020 research and innovation programme under the Marie Sk{\l}odowska-Curie Grant Agreement No 836752 (OncoViroMRI). This paper reflects only the author{\textquoteright}s view and the Research Executive Agency of the European Commission is not responsible for any use that may be made of the information it contains. Publisher Copyright: {\textcopyright} 2020, The Author(s).",
year = "2020",
month = dec,
doi = "10.1038/s41598-020-77576-z",
language = "English (US)",
volume = "10",
journal = "Scientific Reports",
issn = "2045-2322",
publisher = "Nature Publishing Group",
number = "1",
}