Optogenetics and Protein Engineering - The Lin Group
Generation of protein-based tools for neuroscientific and biological research
The goal of the laboratory is to develop novel biochemical and chemical biochemical tools to study fundamental neuroscience questions. These tools can be used by biologists to understand how cellular events lead to behavioural effects in neuroscience, and detect the cellular events that occur during the performance of a behaviour. The laboratory utilises a multi-discipline approach to tackle these scientific questions and we utilise protein-engineering, physiological, optical, molecular biological, chemical and biochemical techniques to develop and assay the tools.
Expertise
- Optogenetic tools
- Chemogenetic tools
- Fluorescent protein based sensors
- Neurobiology of learning and behaviour
Projects
Protein-based actuators for neuroscientific research
Starting with the works conducted within Professor Roger Tsien's laboratory at University of California, San Diego, we have made substantial improvements in existing variants of Channelrhodopsins (ChRs) and developed a novel optogenetic approach to inhibit synaptic release (InSynC). The goals of the current laboratory is to generate more useful tools for neuroscientific and biological researches. Currently we are working with new optogenetic technologies to manipulate intracellular signalling cascade and synaptic plasticity in neurons. When activated with light, we would like to be able to alter the intracellular signalling cascade or synaptic plasticity at a defined temporal window that would enable the manipulation of behaviour in model laboratory organisms. These tools would enable the investigation the neuroscientific question of 'causality' - how is the activities of an individual cell or a particular circuitry lead to the change in behaviour.
Protein-based sensors for neuroscientific research
The works on fluorescent proteins has proved to be an essential part of biological and chemical sciences. These tools have enable us to observe the dynamics of intracellular activities in real-time with high spatial resolution. We are currently undertaking further work to develop fluorescent protein-based sensors that would permit the detection of cellular and synaptic activities. The current focus on the development of sensors for based on the far-red fluorescent protein smURFP that we developed with Dr. Erik Rodriguez
Affiliation
Biomedical Sciences
School of Medicine
Contact
Email: John.lin@utas.edu.au
Group Leader(s)
Group members
- Dr David Gell
- Jenny Smith
- Dr Agnieszka Zbela
- Andrew Reading
- Ann-Sofie Bjerre
Additional Information
Constructs available for request
Lin et al. Biophysical Journal (2009) and Nabavi et al. Nature (2014)
- Mammalian optimized ChD-Citrine (oChD-Citrine) in AAV2 vector (Annotated Genebank file)(Addgene 50976)
- Mammalian optimized ChEF-Citrine (oChEF-Citrine) in AAV2 vector (Annotated Genebank file)(Addgene 50975)
- Mammalian optimized ChIEF-Citrine (oChIEF-Citrine) in AAV2 vector (Annotated Genebank file)(Addgene 50974)
- oChIEF-tdTomato in AAV2 vector (Annotated Genebank file)(Addgene 50977)
- flex-oChIEF-Citrine in AAV2 vector (Annotated Genebank file)(Addgene 50973)
- Maps of AAV2-based vector (AAV2-vectors (PDF 145.8 KB))
Lin et al. Nat Neurosci (2013)
- ReaChR-Citrine in AAV2 vector (Annotated Genebank file) (Addgene 50954)
- Flex-ReaChR-Citrine in AAV2 vector (Annotated Genebank file) (Addgene 50955)
- Maps of the AAV-ReaChR vectors (ReaChR-maps (PDF 43.2 KB))
- ReaChR-Citrine in 2nd generation lentiviral vector (Annotated Genebank file) (Addgene 50956)
- Maps of the pLenti-ReaChR-Citrine vector (pLenti-hSyn-ReaChR-Cit.gb Map (PDF 24.5 KB))
- miniSOG-VAMP2-Citrine in AAV2 vector (Annotated Genebank file) (Addgene 50969)
- miniSOG-VAMP2-T2A-mCherry in AAV2 vector (Annotated Genebank file) (Addgene 50970)
- Synaptophysin (Syp1)-miniSOG-Citrine in AAV2 vector (Addgene 50971)
- Synaptophysin (Syp1)-miniSOG-T2A-mCherry in AAV2 vector (Annotated Genebank file) (Addgene 50972)
- Maps of the InSynC constructs (AAV2-InSynC (PDF 82.8 KB))
Inagaki et al. Nat Methods (2014)
- UAS-ReaChR – Drosophila available at flystocks.bio.indiana.edu (Stock No. 53740-53749)
- Others: UAS-ChIEF – Drosophila available at flystocks.bio.indiana.edu (Stock No. 52228-52230)
Hooks et al. J Neurosci (2015)
- See Dr. Karel Svoboda's Janelia page
Rodriguez et al. Nat Methods (2016)
- See Addgene page
Useful Information / Tools
- The spectral responses of different channelrhodopsin variants (ChR2, ChR2(HR), ChIEF, ReaChR, VChR1, VCOMET, Chrimson and Chronos)(ChR action spectra (XLSX 65.9 KB))
- Fluorescent protein resources – spectral response (Tsien Laboratory Fluorophore Spectra (XLS)), comparison (nic.ucsf.edu/dokuwiki/doku.php?id=fluorescent_proteins, fpvis.org, fpvis.org/PSFP)
- APE plasmid editor - by Wayne Davis and Erik Jorgensen
- UCSF Chimera