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 ([Formula: see text], n = 10).

journal_name

Sci Rep

journal_title

Scientific reports

authors

Perlman O,Ito H,Gilad AA,McMahon MT,Chiocca EA,Nakashima H,Farrar CT

doi

10.1038/s41598-020-77576-z

subject

Has Abstract

pub_date

2020-11-26 00:00:00

pages

20664

issue

1

issn

2045-2322

pii

10.1038/s41598-020-77576-z

journal_volume

10

pub_type

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