Jove
Visualize
Contact Us
JoVE
x logofacebook logolinkedin logoyoutube logo
ABOUT JoVE
OverviewLeadershipBlogJoVE Help Center
AUTHORS
Publishing ProcessEditorial BoardScope & PoliciesPeer ReviewFAQSubmit
LIBRARIANS
TestimonialsSubscriptionsAccessResourcesLibrary Advisory BoardFAQ
RESEARCH
JoVE JournalMethods CollectionsJoVE Encyclopedia of ExperimentsArchive
EDUCATION
JoVE CoreJoVE BusinessJoVE Science EducationJoVE Lab ManualFaculty Resource CenterFaculty Site
Terms & Conditions of Use
Privacy Policy
Policies

Related Concept Videos

Protein Dynamics in Living Cells01:19

Protein Dynamics in Living Cells

2.1K
Different fluorescence-based techniques are used to study the protein dynamics in living cells. These techniques include FRAP, FRET, and PET.
Fluorescent recovery after photobleaching (FRAP) is a fluorescent-protein-based detection technique used to quantify protein movement rates within the cell. This method exposes a small portion of the cell to an intense laser beam. The laser beam causes permanent photobleaching of the fluorophore-tagged proteins in the exposed region. As the bleached...
2.1K
Reporter Genes02:11

Reporter Genes

11.1K
Reporter genes are a type of protein-coding gene that are often tagged to a gene of interest. Once inside a target cell, reporter genes usually produce visually identifiable characteristics like fluorescence and luminescence when expressed along with the gene of interest. Thus, reporter genes “report” the presence or absence of genes of interest in an organism, determine the gene expression pattern, or track the physical location of a DNA segment or protein in the cell.
11.1K

You might also read

Related Articles

Articles linked to this work by shared authors, journal, and citation graph.

Sort by
Same author

Contribution of muscarinic acetylcholine receptors to bottom-up amplification of frontal and parietal cortical responses to rare deviant tones in rats.

Neuroscience·2026
Same author

Discovery of noncanonical cytochrome P450 enzymes in nature.

Nature chemical biology·2026
Same author

Intermittent parathyroid hormone employs autonomous and non-autonomous mechanisms to drive osteogenesis from Ebf3-expressing skeletal progenitor cells.

bioRxiv : the preprint server for biology·2026
Same author

Reply to: A case definition with microscopy would help establish Vibrio as a cause of sea star wasting disease.

Nature ecology & evolution·2026
Same author

Short- and Long-Term Effects of Social Isolation on Adult Murine Bone are Sex-Dependent.

bioRxiv : the preprint server for biology·2026
Same author

Expanding the paradigm of glycopeptide antibiotic recognition through molecular dynamics simulations.

Communications chemistry·2026

Related Experiment Video

Updated: May 24, 2025

Glutamine Flux Imaging Using Genetically Encoded Sensors
10:23

Glutamine Flux Imaging Using Genetically Encoded Sensors

Published on: July 31, 2014

9.5K

State-dependent motion of a genetically encoded fluorescent biosensor.

Paul C Rosen1,2, Samantha M Horwitz3, Daniel J Brooks1

  • 1Department of Neurobiology, Harvard Medical School, Boston, MA 02115.

Proceedings of the National Academy of Sciences of the United States of America
|March 6, 2025
PubMed
Summary

Genetically encoded biosensors like LiLac, used for lactate detection, undergo structural changes. Understanding these dynamics is key to engineering better fluorescent biosensors.

Keywords:
genetically encoded fluorescent biosensormetabolite biosensorprotein conformational change

More Related Videos

Myosin-Specific Adaptations of In vitro Fluorescence Microscopy-Based Motility Assays
08:57

Myosin-Specific Adaptations of In vitro Fluorescence Microscopy-Based Motility Assays

Published on: February 4, 2021

5.8K
Author Spotlight: Advancing Real-Time cAMP Detection in Cells Using cADDis Biosensor
06:03

Author Spotlight: Advancing Real-Time cAMP Detection in Cells Using cADDis Biosensor

Published on: March 22, 2024

795

Related Experiment Videos

Last Updated: May 24, 2025

Glutamine Flux Imaging Using Genetically Encoded Sensors
10:23

Glutamine Flux Imaging Using Genetically Encoded Sensors

Published on: July 31, 2014

9.5K
Myosin-Specific Adaptations of In vitro Fluorescence Microscopy-Based Motility Assays
08:57

Myosin-Specific Adaptations of In vitro Fluorescence Microscopy-Based Motility Assays

Published on: February 4, 2021

5.8K
Author Spotlight: Advancing Real-Time cAMP Detection in Cells Using cADDis Biosensor
06:03

Author Spotlight: Advancing Real-Time cAMP Detection in Cells Using cADDis Biosensor

Published on: March 22, 2024

795

Area of Science:

  • Biochemistry
  • Structural Biology
  • Molecular Imaging

Background:

  • Genetically encoded biosensors offer single-cell resolution for measuring biochemical properties in vivo.
  • Current biosensors function as "black boxes" with limited understanding of their structural states and dynamics.
  • The structural basis for fluorescence changes in biosensors remains largely unexplored.

Purpose of the Study:

  • To elucidate the structural mechanisms underlying the function of the LiLac lactate biosensor.
  • To characterize the low- and high-fluorescence states of LiLac and the transitions between them.
  • To provide insights for the rational design and engineering of novel fluorescent biosensors.

Main Methods:

  • Utilized X-ray crystallography to determine the three-dimensional structures of LiLac in different states.
  • Employed engineered high-affinity metal bridges to probe structural dynamics.
  • Leveraged quantitative fluorescence-lifetime imaging to correlate structural changes with sensor output.

Main Results:

  • LiLac exhibits a significant interdomain twist motion correlating with lactate binding.
  • The high-lifetime state (low lactate) corresponds to a "sealed" conformation, while the low-lifetime state (high lactate) adopts a "cracked" conformation.
  • Structural plasticity and interdomain dynamics are critical for LiLac's fluorescence response.

Conclusions:

  • The study reveals the dynamic structural rearrangements in LiLac that enable lactate detection.
  • Understanding these structure-dynamics relationships is crucial for advancing the field of genetically encoded biosensors.
  • These findings provide a foundation for engineering improved biosensors with tailored properties.