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13 C MR reporter probe system using dynamic nuclear polarization
Author(s) -
Chen Albert P.,
Hurd Ralph E.,
Gu Yiping,
Wilson David M.,
Cunningham Charles H.
Publication year - 2011
Publication title -
nmr in biomedicine
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.278
H-Index - 114
eISSN - 1099-1492
pISSN - 0952-3480
DOI - 10.1002/nbm.1618
Subject(s) - reporter gene , chemistry , transfection , acetylation , hek 293 cells , in vivo , enzyme , microbiology and biotechnology , nuclear magnetic resonance , biochemistry , gene expression , gene , biology , physics
Reporter‐based cell detection and localization in vivo may become an important imaging tool with the emergence of cellular therapy. With the strong signal enhancement provided by dynamic nuclear polarization, an NMR‐based reporter probe system utilizing specific enzyme expression and activity can potentially provide stable, high‐resolution visualization of the cells of interest noninvasively. In this work, a proof‐of‐concept 13 C MR reporter system, using the aminoacylase‐1 reporter gene ( Acy‐1 ) and prepolarized [1‐ 13 C] N ‐acetyl‐ L ‐methionine as the paired substrate, was developed. Using a 3‐T MR scanner, the feasibility of detecting and imaging de‐acetylation of the prepolarized 13 C‐labeled substrate by the aminoacylase‐1 enzyme was demonstrated with purified protein in solution by dynamic 13 C MRS and two‐dimensional MRSI experiments. The potential to perform targeted MRI of cells using this system was also demonstrated by 13 C MR measurement of aminoacylase‐1 activity in HEK 293 cells transfected with the Acy‐1 gene. The de‐acetylation of the substrate was not observed in control cells. Copyright © 2010 John Wiley & Sons, Ltd.