A Vertical Comet Assay for Measuring DNA Damage to Radiation

Space radiation detrimentally affects biological systems and individual organisms. Earth’s magnetic field provides protection to terrestrial inhabitants. However, space missions beyond the low Earth orbit poses increased risk to the health of humans because of radiation in space. Both components of space radiation, chronic exposure to low-dose to galactic cosmic rays and random short-term exposures to the energetic particles from the Sun, might increase risk of malignant transformation. Damage to the genetic material of cells, its DNA, is considered to be the first and most important step in the malignant transformation of normal cell into cancerous one. As this concern is addressed in NASA’s 2012 roadmap in the domain of Human Health, Life Support and Habitation Systems (HLHS), however, currently we do not have reliable techniques to measure DNA damage on a board of spacecraft. A reliable and sensitive technique to collect data at ground and in flight for radiation risk assessment is in demand by NASA. When exposed to radiation, single and double strand-breaks in cellular DNA are produced. The amount of strand-breaks can therefore be correlated to the extent of damage to cells under radiation. One of the most convenient models to study DNA damage and repair process in the live cells is the comet assay, or single cell gel electrophoresis technique. This technique is currently configured to conduct a measurement on live cells under laboratory set-ups. Therefore, quite often the existing comet assay is not suitable for “in field” applications. Development of a simple and sensitive technique would allow researchers to study crucial cellular processes outside of laboratory boundaries and open possibilities for evaluating many environmental exposures currently unobtainable, such as DNA damage during short space flight or long-term interplanetary explorations within the scope of NASA’s missions. In this project we are aimed at developing a new technique, we call it vertical comet or v-comet, to measure the level of DNA damage in live cells or fix-preserved cellular DNA samples. This new assay will be prototyped and tested using the microfluidics technique. We will (a) develop a microchamber containing live cells or preserved cellular DNA; (b) use electrophoresis to sample damaged DNA by pulling broken strands into a circulating buffer in a closed-loop; (c) electrophoresis is continuously applied while the buffer is circulating to accumulate the broken strands; and (d) the amount of DNA will be measured by a sensitive fluorescent labeling. By taking the advantages of small scales of microchamber configuration, in this proposal we will design and test an integrated approach combining sensitivity of traditional comet-assay technique with the versatility and subtle precision of microfluidics technique. We envision that this new technique will allow researchers to measure DNA damage in the field conditions, such as .to help astronauts avoid overexposure to space radiation, and construct a correlation between the doses of chronic space radiation and the extent of cumulative DNA damage. This project explicitly addresses NASA missions within the area of space radiation, as evidenced by two letters of support from NASA: the Space Biosciences Branch at the NASA Ames Research Center, and the Radiation Biophysics Laboratory at Johnson Space Center. This project will enhance the research infrastructure by engaging faculty and students crossing the disciplines at the University of Alaska Fairbanks. The project will be self-sustained by developing a few proposals to NSF and NIH during and after the project phase. The innovation of a v-comet assay is on either live cells or sampled cells in a conserved, dried platform to study DNA damage not bounded by the laboratory environment. Commercialization will be pursued for licensing and patenting the proposed technique.

This is a mature technology (TRL 7+) — the realistic path in is usually NASA's Technology Transfer Program: licensing an existing NASA patent, or a Space Act Agreement to use NASA facilities/expertise directly. NASA also runs a startup licensing program with no upfront fee for companies formed to commercialize a specific NASA technology.

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