Directed Molecular Evolution Reveals Gaussia Luciferase Variants with Enhanced Light Output Stability

被引：35

作者：

Degeling, M. Hannah ^{[1
,2
,3
]}

Bovenberg, M. Sarah S. ^{[1
,2
,3
]}

Lewandrowski, Grant K. ^{[1
,2
]}

de Gooijer, Mark C. ^{[1
,2
,4
]}

Vleggeert-Lankamp, Carmen L. A. ^{[3
]}

Tannous, Marie ^{[5
]}

Maguire, Casey A. ^{[1
,2
]}

Tannous, Bakhos A. ^{[1
,2
]}

机构：

[1] Massachusetts Gen Hosp, Ctr Neurosci, Expt Therapeut & Mol Imaging Lab, Dept Neurol, Charlestown, MA 02129 USA

[2] Harvard Univ, Sch Med, Program Neurosci, Boston, MA 02114 USA

[3] Leiden Univ, Med Ctr, Dept Neurosurg, Leiden, Netherlands

[4] Canc Ctr Amsterdam, VU Med Ctr, Dept Neurosurg, Neurooncol Res Grp, NL-1007 MB Amsterdam, Netherlands

[5] Notre Dame Univ, Fac Nat & Appl Sci, Barsa, Lebanon

来源：

ANALYTICAL CHEMISTRY | 2013年 / 85卷 / 05期

关键词：

REPORTER GENE-EXPRESSION; BIOLUMINESCENCE;

D O I：

10.1021/ac4003134

中图分类号：

O65 [分析化学];

学科分类号：

070302 ; 081704 ;

摘要：

Gaussia Luciferase (Gluc) has proven to be a powerful mammalian cell reporter for monitoring numerous biological processes in immunology, virology, oncology, and neuroscience. Current limitations of Gluc as a reporter include its emission of blue light, which is absorbed by mammalian tissues, limiting its use in vivo, and a flash-type bioluminescence reaction, making it unsuited for high-throughput applications. To overcome these limitations, a library of Gluc variants was generated using directed molecular evolution and screened for relative light: output, a shift in emission spectrum, and glow-type light emission kinetics. Several variants with a 10-15 nm shift in their light emission peak were found. Further, a Gluc variant that catalyzes a glow-type bioluminescence reaction, suited for high-throughput applications, was also identified. These results indicate that molecular evolution could be used to modulate Gluc bioluminescence reaction characteristics.

引用

页码：3006 / 3012

页数：7

共 23 条

[1] Bioluminescence imaging: progress and applications [J].

Badr, Christian E. ;

Tannous, Bakhos A. .

TRENDS IN BIOTECHNOLOGY, 2011, 29 (12) :624-633

[2] Functional Drug Screening Assay Reveals Potential Glioma Therapeutics [J].

Badr, Christian E. ;

Wurdinger, Thomas ;

Tannous, Bakhos A. .

ASSAY AND DRUG DEVELOPMENT TECHNOLOGIES, 2011, 9 (03) :281-289

[3] Tumor-specific imaging through progression elevated gene-3 promoter-driven gene expression [J].

Bhang, Hyo-eun C. ;

Gabrielson, Kathleen L. ;

Laterra, John ;

Fisher, Paul B. ;

Pomper, Martin G. .

NATURE MEDICINE, 2011, 17 (01) :123-U302

[4] Optical imaging of Renilla luciferase reporter gene expression in living mice [J].

Bhaumik, S ;

Gambhir, SS .

PROCEEDINGS OF THE NATIONAL ACADEMY OF SCIENCES OF THE UNITED STATES OF AMERICA, 2002, 99 (01) :377-382

[5] Enhanced Gaussia Luciferase Blood Assay for Monitoring of in Vivo Biological Processes [J].

Bovenberg, M. Sarah S. ;

Degeling, M. Hannah ;

Tannous, Bakhos A. .

ANALYTICAL CHEMISTRY, 2012, 84 (02) :1189-1192

[6] Multi-parameter phenotypic profiling: using cellular effects to characterize small-molecule compounds [J].

Feng, Yan ;

Mitchison, Timothy J. ;

Bender, Andreas ;

Young, Daniel W. ;

Tallarico, John A. .

NATURE REVIEWS DRUG DISCOVERY, 2009, 8 (07) :567-578

[7] Identification of two catalytic domains in a luciferase secreted by the copepod Gaussia princeps [J].

Inouye, Satoshi ;

Sahara, Yuiko .

BIOCHEMICAL AND BIOPHYSICAL RESEARCH COMMUNICATIONS, 2008, 365 (01) :96-101

[8] Two-step transcriptional amplification as a method for imaging reporter gene expression using weak promoters [J].

Iyer, M ;

Wu, L ;

Carey, M ;

Wang, Y ;

Smallwood, A ;

Gambhir, SS .

PROCEEDINGS OF THE NATIONAL ACADEMY OF SCIENCES OF THE UNITED STATES OF AMERICA, 2001, 98 (25) :14595-14600

[9] Superluminescent Variants of Marine Luciferases for Bioassays [J].

Kim, Sung Bae ;

Suzuki, Hideyuki ;

Sato, Moritoshi ;

Tao, Hiroaki .

ANALYTICAL CHEMISTRY, 2011, 83 (22) :8732-8740

[10] Gaussia Luciferase Variant for High-Throughput Functional Screening Applications [J].

Maguire, Casey A. ;

Deliolanis, Nikolaos C. ;

Pike, Lisa ;

Niers, Johanna M. ;

Tjon-Kon-Fat, Lee-Ann ;

Sena-Esteves, Miguel ;

Tannous, Bakhos A. .

ANALYTICAL CHEMISTRY, 2009, 81 (16) :7102-7106

← 1 2 3 →