RT Journal Article
SR Electronic
T1 An Engineered Amber-Emitting Nano Luciferase and Its Use for Immunobioluminescence Imaging in Vivo
JF bioRxiv
FD Cold Spring Harbor Laboratory
SP 2022.05.23.493143
DO 10.1101/2022.05.23.493143
A1 Ying Xiong
A1 Yiyu Zhang
A1 Zefan Li
A1 Md Shamim Reza
A1 Xinyu Li
A1 Xiaodong Tian
A1 Huiwang Ai
YR 2022
UL http://biorxiv.org/content/early/2022/05/24/2022.05.23.493143.abstract
AB The NanoLuc luciferase (NLuc) and its furimazine (FRZ) substrate have revolutionized bioluminescence (BL) assays and imaging. However, the use of the NLuc-FRZ luciferase-luciferin pair for mammalian tissue imaging is hindered by the low tissue penetration of the emitting blue photons. Here, we present the development of an NLuc mutant, QLuc, which catalyzes the oxidation of a synthetic QTZ luciferin for bright and red-shifted emission peaking at ∼ 585 nm. This amber-light-emitting luciferase-luciferin pair exhibited improved performance for imaging deep-tissue targets in live mice. Leveraging this novel bioluminescent reporter, we further pursued in vivo immunobioluminescence imaging (immunoBLI), which used a fusion protein of a single-chain variable antibody fragment (scFv) and QLuc for molecular imaging of tumor-associated antigens in a xenograft mouse model. As one of the most red-shifted NLuc variants, we expect QLuc to find broad applications in noninvasive imaging in mammals. Moreover, the immunoBLI method complements immunofluorescence imaging and immuno-positron emission tomography (immunoPET), serving as a convenient and nonradioactive molecular imaging tool for animal models in basic and preclinical research.Competing Interest StatementHA and YX are listed as inventors of a patent or a patent application covering some luciferase and luciferin variants described in this work.