Novel NanoLuc substrates enable bright two-population bioluminescence imaging in animals

Su, Yichi; Walker, Joel R.; Park, Yun‐Hee; Smith, Thomas P.; Liu, Lan Xiang; Hall, Mary P.; Labanieh, Louai; Hurst, Robin; Wang, David C.; Encell, Lance P.; Kim, Namdoo; Zhang, Feijie; Kay, Mark A.; Casey, Kerriann M.; Majzner, Robbie G.; Cochran, Jennifer R.; Mackall, Crystal L.; Kirkland, Thomas A.; Lin, Michael Z.

doi:10.1038/s41592-020-0889-6

Cited by 153 publications

(219 citation statements)

References 32 publications

Supporting

Mentioning

203

Contrasting

Order By: Relevance

“…With complete elimination of any brain implant and head tethering owing to the deep-brain penetration of NIR-II photons, our approach could afford wide applications in dissecting the complex neural circuits of normally behaving animals in a naturally interacting environment such as the IntelliCage 48 . When combined with a red-shifted bioluminescent reporter for noninvasive neural activity imaging 57 , our method may realize an all-optical bidirectional neural interface through the scalp and skull in behaving animals.…”

Section: Discussionmentioning

confidence: 99%

Through-scalp deep-brain stimulation in tether-free, naturally-behaving mice with widefield NIR-II illumination

Jiang

Rommelfanger

et al. 2020

Preprint

View full text Add to dashboard Cite

Neural modulation techniques with electricity, light and other forms of energy have enabled the deconstruction of neural circuitry. One major challenge of existing neural modulation techniques is the invasive brain implants and the permanent skull attachment of an optical fiber for modulating neural activity in the deep brain. Here we report an implant-free and tether-free optical neuromodulation technique in deep-brain regions through the intact scalp with brain-penetrant second near-infrared (NIR-II) illumination. Macromolecular infrared nanotransducers for deep-brain stimulation (MINDS) demonstrate exceptional photothermal conversion efficiency of 71% at 1064 nm, the wavelength that minimizes light attenuation by the brain in the entire 400-1700 nm spectrum. Upon widefield 1064-nm illumination >50 cm above the mouse head at a low incident power density of 10 mW/mm2, deep-brain neurons are activated by MINDS-sensitized TRPV1 channels with minimal thermal damage. Our approach could open opportunities for simultaneous neuromodulation of multiple socially interacting animals by remotely irradiating NIR-II light to stimulate each subject individually.

show abstract

Section: Discussionmentioning

confidence: 99%

Through-scalp deep-brain stimulation in tether-free, naturally-behaving mice with widefield NIR-II illumination

Jiang

Rommelfanger

et al. 2020

Preprint

View full text Add to dashboard Cite

show abstract

“…We anticipate that pre-clinical research into (oncolytic) viral infections will strongly benefit from this non-invasive bioluminescent viral screening tool, enabling fast and simple longitudinal readout protocols applicable for rapid drug screening and viral-risk virus infectivity and transduction in a range of cell lines. To enable in vivo imaging of the infection course with our HiBiT-reporter virus, we employed hydrofurimazine, a substrate with improved solubility compared to furimazine [13]. The substrate was formulated this in a non-toxic, highly water-soluble excipient, which allows much higher substrate loading and extended light emission in vivo after a single substrate injection.…”

Section: Discussionmentioning

confidence: 99%

“…The HFz substrate in its novel formulation proved particularly well-suited for these experiments, where a broad imaging time window was required. In fact, the substrate showed a stable signal 15 min after intraperitoneal administration when formulated in P-407 in mice carrying NanoLuc expressing cells [13]. As early as 24 h post viral injection, all mice showed specific photon emission, suggesting successful infection of PC-3-LgBiT tumor cells in vivo and good distribution of HFz to the tumor site after i.p.…”

Section: Sensitive Longitudinal In Vivo Monitoring Of Hibit-reporter mentioning

confidence: 92%

See 1 more Smart Citation

NanoBiT System and Hydrofurimazine for Optimized Detection of Viral Infection in Mice—A Novel in Vivo Imaging Platform

Gaspar

Zambito

Dautzenberg

et al. 2020

IJMS

Self Cite

View full text Add to dashboard Cite

Reporter genes are used to visualize intracellular biological phenomena, including viral infection. Here we demonstrate bioluminescent imaging of viral infection using the NanoBiT system in combination with intraperitoneal injection of a furimazine analogue, hydrofurimazine. This recently developed substrate has enhanced aqueous solubility allowing delivery of higher doses for in vivo imaging. The small high-affinity peptide tag (HiBiT), which is only 11 amino-acids in length, was engineered into a clinically used oncolytic adenovirus, and the complementary large protein (LgBiT) was constitutively expressed in tumor cells. Infection of the LgBiT expressing cells with the HiBiT oncolytic virus will reconstitute NanoLuc in the cytosol of the cell, providing strong bioluminescence upon treatment with substrate. This new bioluminescent system served as an early stage quantitative viral transduction reporter in vitro and also in vivo in mice, for longitudinal monitoring of oncolytic viral persistence in infected tumor cells. This platform provides novel opportunities for studying the biology of viruses in animal models.

show abstract

“…Of note, other efforts to improve the sensitivity of BLI for in vivo imaging include directed evolution of luciferase from marine organisms, which typically emit blue light, such as Nanoluc, for alternative substrates that emit light of longer wave lengths, or the coupling of blue-emitting BLI systems with orange fluorescent proteins for FRET effects. 12 , 13 Such approaches may potentially reach in vivo sensitivity similar to that of Akaluc BLI and represent useful additions due to their distinct substrate specificities, thus allowing 2-population BLI. 13 …”

Section: Discussionmentioning

confidence: 99%

Akaluc bioluminescence offers superior sensitivity to track in vivo glioma expansion

Bozec

Sattiraju

Bouras

et al. 2020

Neuro-Oncology Advances

View full text Add to dashboard Cite

Background Longitudinal tracking of tumor growth using non-invasive bioluminescence imaging (BLI) is a key approach for studies of in vivo cancer models, with particular relevance for investigations of malignant gliomas in rodent intracranial transplant paradigms. Akaluciferase (Akaluc) is a new BLI system with higher signal strength than standard firefly luciferase (Fluc). Here, we establish Akaluc BLI as a sensitive method for in vivo tracking of glioma expansion. Methods We engineered a lentiviral vector for expression of Akaluc in high-grade glioma cell lines, including patient-derived glioma stem cell (GSC) lines. Akaluc expressing glioma cells were compared to matching cells expressing Fluc in both in vitro and in vivo BLI assays. We also conducted proof-of-principle BLI studies with intracranial transplant cohorts receiving chemo-radiation therapy. Results Akaluc-expressing glioma cells produced more than 10 times higher BLI signals than Fluc-expressing counterparts when examined in vitro, and more than 100-fold higher signals when compared to Fluc-expressing counterparts in intracranial transplant models in vivo. The high sensitivity of Akaluc permitted detection of intracranial glioma transplants starting as early as 4 hours after implantation and with as little as 5,000 transplanted cells. The sensitivity of the system allowed us to follow engraftment and expansion of intracranial transplants of GSC lines. Akaluc was also robust for sensitive detection of in vivo tumor regression after therapy and subsequent relapse. Conclusion Akaluc BLI offers superior sensitivity for in vivo tracking of glioma in the intracranial transplant paradigm, facilitating sensitive approaches for the study of glioma growth and response to therapy.

show abstract

Novel NanoLuc substrates enable bright two-population bioluminescence imaging in animals

Cited by 153 publications

References 32 publications

Through-scalp deep-brain stimulation in tether-free, naturally-behaving mice with widefield NIR-II illumination

Through-scalp deep-brain stimulation in tether-free, naturally-behaving mice with widefield NIR-II illumination

NanoBiT System and Hydrofurimazine for Optimized Detection of Viral Infection in Mice—A Novel in Vivo Imaging Platform

Akaluc bioluminescence offers superior sensitivity to track in vivo glioma expansion

Contact Info

Product

Resources

About