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Am J Transl Res 2011;3(4):392-403
Original Article
HaloTag: a novel reporter gene for positron emission tomography
Hao Hong, Hélène A. Benink, Yin Zhang, Yunan Yang, H. Tetsuo Uyeda, Jonathan W. Engle, Gregory W. Severin,
Mark G. McDougall, Todd E. Barnhart, Dieter H. Klaubert, Robert J. Nickles, Frank Fan, Weibo Cai
Department of Radiology, University of Wisconsin - Madison, Madison, WI, USA; Promega Corporation, Madison,
WI, USA; Department of Medical Physics, University of Wisconsin - Madison, Madison, WI, USA; Promega
Biosciences, LLC, San Luis Obispo, CA, USA; University of Wisconsin Carbone Cancer Center, Madison, WI, USA.
Received July 12, 2011; accepted July, 2011; Epub July, 2011; Published July, 2011
Abstract: Among the many molecular imaging techniques, reporter gene imaging has been a dynamic area of
research. The HaloTag protein is a modified haloalkane dehalogenase which was designed to covalently bind to
synthetic ligands (i.e. the HaloTag ligands [HTL]). Covalent bond formation between the HaloTag protein and the
chloroalkane within the HTL occurs rapidly under physiological conditions, which is highly specific and essentially
irreversible. Over the years, HaloTag technology has been investigated for various applications such as in vitro/in
vivo imaging, protein purification/trafficking, high-throughput assays, among others. The goal of this study is to
explore the use of the HaloTag protein as a novel reporter gene for positron emission tomography (PET) imaging.
By attaching a HaloTag-reactive chloroalkane to 1, 4, 7-triazacyclononane-N, N', N''-triacetic acid (NOTA) through
hydrophilic linkers, the resulting NOTA-conjugated HTLs were labeled with 64Cu and tested for PET imaging in
living mice bearing 4T1-HaloTag-ECS tumors, which stably express the HaloTag protein on the cell surface.
Significantly higher uptake of 64Cu-NOTA-HTL-S (which contains a short hydrophilic linker) in the
4T1-HaloTag-ECS than the non-HaloTag-expressing 4T1 tumors was observed, which demonstrated the
HaloTag specificity of 64Cu-NOTA-HTL-S and warranted future investigation of the HaloTag protein as a PET
reporter gene.(AJTR1107002).
Keywords: Reporter gene, HaloTag, positron emission tomography (PET), cell tracking, cancer, molecular
imaging
Full Text PDF
Address all correspondence to:
Weibo Cai, PhD
Departments of Radiology and Medical Physics
University of Wisconsin - Madison
Madison, WI 53705-2275, USA
Phone: 608-262-1749; Fax: 608-265-0614
Email: wcai@uwhealth.org
Original Article
HaloTag: a novel reporter gene for positron emission tomography
Hao Hong, Hélène A. Benink, Yin Zhang, Yunan Yang, H. Tetsuo Uyeda, Jonathan W. Engle, Gregory W. Severin,
Mark G. McDougall, Todd E. Barnhart, Dieter H. Klaubert, Robert J. Nickles, Frank Fan, Weibo Cai
Department of Radiology, University of Wisconsin - Madison, Madison, WI, USA; Promega Corporation, Madison,
WI, USA; Department of Medical Physics, University of Wisconsin - Madison, Madison, WI, USA; Promega
Biosciences, LLC, San Luis Obispo, CA, USA; University of Wisconsin Carbone Cancer Center, Madison, WI, USA.
Received July 12, 2011; accepted July, 2011; Epub July, 2011; Published July, 2011
Abstract: Among the many molecular imaging techniques, reporter gene imaging has been a dynamic area of
research. The HaloTag protein is a modified haloalkane dehalogenase which was designed to covalently bind to
synthetic ligands (i.e. the HaloTag ligands [HTL]). Covalent bond formation between the HaloTag protein and the
chloroalkane within the HTL occurs rapidly under physiological conditions, which is highly specific and essentially
irreversible. Over the years, HaloTag technology has been investigated for various applications such as in vitro/in
vivo imaging, protein purification/trafficking, high-throughput assays, among others. The goal of this study is to
explore the use of the HaloTag protein as a novel reporter gene for positron emission tomography (PET) imaging.
By attaching a HaloTag-reactive chloroalkane to 1, 4, 7-triazacyclononane-N, N', N''-triacetic acid (NOTA) through
hydrophilic linkers, the resulting NOTA-conjugated HTLs were labeled with 64Cu and tested for PET imaging in
living mice bearing 4T1-HaloTag-ECS tumors, which stably express the HaloTag protein on the cell surface.
Significantly higher uptake of 64Cu-NOTA-HTL-S (which contains a short hydrophilic linker) in the
4T1-HaloTag-ECS than the non-HaloTag-expressing 4T1 tumors was observed, which demonstrated the
HaloTag specificity of 64Cu-NOTA-HTL-S and warranted future investigation of the HaloTag protein as a PET
reporter gene.(AJTR1107002).
Keywords: Reporter gene, HaloTag, positron emission tomography (PET), cell tracking, cancer, molecular
imaging
Full Text PDF
Address all correspondence to:
Weibo Cai, PhD
Departments of Radiology and Medical Physics
University of Wisconsin - Madison
Madison, WI 53705-2275, USA
Phone: 608-262-1749; Fax: 608-265-0614
Email: wcai@uwhealth.org
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