Mechanistic analysis of gene-expression machinery and DNA nanodevices using single-molecule fluorescence spectroscopy
Lead Research Organisation:
University of Oxford
Department Name: Oxford Physics
Abstract
This proposal describes the construction and further development of a special, ultra-sensitive light microscope along with proposals for using this microscope to study how tiny biological machines work and how DNA (the basis of genetic material) can be used to transport electricity or energy.The microscope, which we refer to as a single-molecule fluorescence microscope , is specifically designed to allow detection and monitoring of individual ( single ) fluorescent molecules present in a detection zone (as opposed to conventional microscopes that require thousands or millions of molecules to be present in a detection zone). An additional unique feature of the microscope rests on the fact that two or three lasers are used in rapid alternation (alternating much faster than the average time that one molecule spends in the detection zone) for the detection and the analysis of single molecules; the alternating-laser microscope format carries several advantages compared to earlier single-laser versions. This version of the single-molecule fluorescence microscope allows us to determine the number of parts that make up a biological machine, to measure how strong the parts bind to each other, how far apart the parts are spaced and at what orientation, and what are the movements of the parts when the tiny biological machine works.We will use this special microscope to understand processes that occur during gene expression, the path that leads from the genetic information (stored in DNA) to the manufacturing of proteins (the molecules that make up most of the machines and structures of living cells). Specifically, we will focus on the process of gene transcription, which uses tiny biological machines that read DNA and copy the information into a messenger molecule (messenger RNA). We will also study the process of gene silencing, which uses tiny biological machines that recognise and destroy messenger RNA. Finally, we will use our special microscope to develop strategies that will allow us to use DNA molecules as tiny wires that conduct electricity and energy; these DNA-based wires can be useful in building smaller and more efficient electronic circuits.Our results will help the scientific community to understand how gene expression works and why certain diseases arise when gene expression malfunctions; our work on gene expression will also aid in the development of new drugs that will improve health. Finally, our work on DNA-based wires will be helpful in developing the next generation of electronics.
People |
ORCID iD |
Achillefs Kapanidis (Principal Investigator) |
Publications
Goodman RP
(2009)
A facile method for reversibly linking a recombinant protein to DNA.
in Chembiochem : a European journal of chemical biology
Doose S
(2007)
Periodic acceptor excitation spectroscopy of single molecules.
in European biophysics journal : EBJ
Sustarsic M
(2014)
Optimized delivery of fluorescently labeled proteins in live bacteria using electroporation.
in Histochemistry and cell biology
Goodman RP
(2008)
Reconfigurable, braced, three-dimensional DNA nanostructures.
in Nature nanotechnology
Kapanidis AN
(2009)
Biology, one molecule at a time.
in Trends in biochemical sciences
N/a Heilemann
(2007)
Single-molecule studies of sigma54-dependent transcription
Description | - Construction and characterization of three-laser single-molecule fluorescence microscope for studies of genetic processes and DNA nanodevices - Development of single-molecule assays for studying sigma-54 dependent transcription - Purification of human Argonaute 2 and development of protocols for fluorescent labeling of RNA - Development of assays of electron and energy-transfer on short DNA molecules |
Exploitation Route | Microscopes and single-molecule assays and biosensors for use in the university education (student training), the biomedical sector (biosensors, microbe detection), biodefence (pathogen detection), food industry (pathogen detection), animal health. - Subsequent work focused on construction of small microscopes for use in the academic and biomedical environment |
Sectors | Agriculture, Food and Drink,Education,Pharmaceuticals and Medical Biotechnology,Security and Diplomacy |
URL | http://www.physics.ox.ac.uk/users/kapanidis/Group/Main.ResearchInterests.html |
Description | Imperial College London |
Organisation | Imperial College London |
Country | United Kingdom |
Sector | Academic/University |
Start Year | 2006 |
Title | DNA-BASED BIOSENSORS |
Description | There is described a method of detection of the protein-dependent coincidence of DNA in a sample which comprises detection using luminescence of one or more luminophores introduced into DNA with one, two or more DNA fragments which fragments are bound using one or more DNA-binding proteins. |
IP Reference | WO2008099163 |
Protection | Patent application published |
Year Protection Granted | 2008 |
Licensed | No |
Impact | Led to funding that supported further development of instrumentation and biosensors. The instrumentation led to additional IP and to a spin-out company that now employs 100 people. |