E-Z Guide to LMO3

What is BL-OG? Chemogenetic and/or Optogenetic Control in a Single Molecule

Schematic of Basic BL-OG

In BioLuminescent OptoGenetics (BL-OG), molecular light drives an optogenetic sensor. LuMinOpsins (LMOs) are single molecules that tether the light producer (luciferase) and opsin. In LMO3, bioluminescent (BL) light drives VChR1 causing depolarization when coelenterazine (CTZ) is present. LMO3 has now been vetted in multiple cell types and contexts.

Why BL-OG? More Flexible and More Trackable = More Useful

Systemic impact (chemogenetic) and specific control (optogenetic) in the same mouse: Study I

Bioluminescent light reports the timing of neuromodulation after injection: Study II

Choose the timing; fast optogenetic and multiple chemogenetic time courses: Study II

Long-term biocompatibility: Studies I-IV

How to Get BL-OG

Order LMO3s as pAAV plasmids from Addgene (114099, 114103, 114104, 114105)

Order native CTZ from Nanolight

The LMO3-floxed mouse is available at Jackson Laboratory (Stock Number: 034853)

How to Use BL-OG

Read more, including a step by step protocol, and download a materials list: Crespo EL, Björefeldt A, Prakash M, Hochgeschwender U. 2021. Bioluminescent optogenetics 2.0: harnessing bioluminescence to activate photosensory proteins in vitro and in vivo. J Vis Exp (174).

Help with BL-OG

Our in-house workshops train users in all aspects of BL-OG use, and our emissaries can provide hands-on training in your lab. Email us for more information.

Toxicity? No. Brain Permeability? Yes.

Toxicology screens commissioned by Prolume (ms in prep., H. Schmidt) did not reveal any toxicity concerns of CTZ, the luciferin we employ, consistent with our extensive experience with repeated administration of it in water-soluble form. Another FAQ is whether CTZ can reach brain targets. Multiple published studies and our ongoing work show that CTZ can access the brain (e.g., neocortex, Choroid Plexus) after peripheral injection.

LMO3: In vivo Studies

Study I Figure

Study I: Impacting Development with Chemogenetic LMO3 Activation, then Probing Changes in the Adult with Optogenetic Drive

Medendorp et al. 2021 drove pyramidal neurons expressing LMO3 by IP CTZ injection P4-P10. They then used direct optogenetic drive of LMO3 to test adult excitability in the same cells. Adult mice showed robust behavioral and physiological changes after developmental BL-OG treatment.

Study II Figure

Study II: Bioluminescent Light from LMO3 Reports Chemogenetic Activation in the Choroid Plexus

Shipley et al. (in review) describes novel imaging and control tools for in vivo Choroid Plexus (ChP) study. LMO3 activation in ChP showed distinct time courses for different CTZ injection routes.

Study III Figure

Study III: IPSC control by LMO3 Post-Stroke: Chemogenetic or Optogenetic Activation Drives Behavioral Recovery and Synaptic Enhancement

Yu et al. (2019 J Neurosci) expressed LMO3 in induced pluripotent stem cells (IPSCs) implanted in vivo post-stroke. Chemogenetic or optogenetic LMO3 activation improved behavioral and sensory responses in vivo, and enhanced peri-infarct synaptic plasticity.

Study IV Figure

Study IV: LMO3 Activation Drives Motor Recovery Following Spinal Cord Injury

Petersen et al. (2022) expressed LMO3 in the ventral cord below an impact injury. Driving LMO3 activation with repeated CTZ injections (10 days) significantly enhanced the long-term recovery of motor coordination/control of the lower limbs, as compared to vehicle control treatment of injured mice.

Questions? Ready to get started with LMO3? Email us!

Click here for a printer-friendly version of our E-Z Guide.

Contact Us