Redesigned reporter gene for improved proton exchange-based molecular MRI contrast.
Redesigned reporter gene for improved proton exchange-based molecular MRI contrast.
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DOI：
10.1038/s41598-020-77576-z
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发表时间：
2020-11-26
影响因子：
4.6
通讯作者：
Farrar CT
中科院分区：
文献类型：
--
作者：
Perlman O;Ito H;Gilad AA;McMahon MT;Chiocca EA;Nakashima H;Farrar CT
报告基因成像允许对活细胞中的分子过程进行非侵入性监测，为病理和治疗的潜在机制提供见解。富赖氨酸蛋白（LRP）化学交换饱和转移（CEST）MRI报告基因先前已被开发并用于肿瘤细胞成像，心脏病毒基因转移和溶瘤病毒治疗。然而，LRP报告基因序列的高度重复性质导致DNA重组事件和一系列截短的LRP蛋白片段的表达，从而极大地限制了CEST灵敏度。在这里，我们报告使用重新设计的LRP报告（rdLRP），旨在提供出色的稳定性和CEST灵敏度。rdLRP不含DNA重复序列或富含GC的区域，带正电荷的氨基酸少30%。用rdLRP转染的细胞裂解物的RT-PCR证明了具有对应于全长DNA的单一独特条带的稳定报告基因。在rdLRP转染细胞的细胞裂解物和体内表达LRP的小鼠脑肿瘤中获得CEST-MRI对比度的明显增加（n = 10）。
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 (, n = 10).
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影响因子：
4
作者：
Bar-Shir, Amnon;Liu, Guanshu;Chan, Kannie W. Y.;Oskolkov, Nikita;Song, Xiaolei;Yadav, Nirbhay N.;Walczak, Piotr;McMahon, Michael T.;van Zijl, Peter C. M.;Bulte, Jeff W. M.;Gilad, Assaf A.
通讯作者：
Gilad, Assaf A.
影响因子：
41.2
作者：
Lu GJ;Farhadi A;Szablowski JO;Lee-Gosselin A;Barnes SR;Lakshmanan A;Bourdeau RW;Shapiro MG
通讯作者：
Shapiro MG
DOI：
10.1152/ajpheart.00935.2010
发表时间：
2011-06-01
影响因子：
4.8
作者：
Campan, Manuela;Lionetti, Vincenzo;Recchia, Fabio A.
通讯作者：
Recchia, Fabio A.
影响因子：
2.9
作者：
Airan, Raag D.;Li, Nan;Pelled, Galit
通讯作者：
Pelled, Galit
影响因子：
5.1
作者：
Chaturvedi P;Zhao B;Zimmerman DL;Belmont AS
通讯作者：
Belmont AS