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The RABiT: high-throughput technology for assessing global DSB repair.

Helen C Turner¹, P Sharma, J R Perrier

¹Department of Radiation Oncology, Center for Radiological Research, Columbia University Medical Center, 630 W. 168th St. VC11-240, New York, NY, 10032, USA, ht2231@columbia.edu.

Radiation and Environmental Biophysics

|January 31, 2014

Summary

The Rapid Automated Biodosimetry Tool (RABiT) offers high-throughput radiation biodosimetry using fingerstick blood. This study characterizes DNA repair kinetics in lymphocytes, demonstrating RABiT

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Area of Science:

Radiation biology
Biotechnology
Molecular biology

Background:

Large-scale radiological events necessitate rapid and accurate radiation biodosimetry.
The Rapid Automated Biodosimetry Tool (RABiT) is an automated, ultra-high-throughput workstation for biodosimetry using fingerstick blood samples.
RABiT is being adapted for laboratory settings to support epidemiological and clinical studies, with a goal to measure DNA repair protein kinetics.

Purpose of the Study:

To characterize the DNA repair kinetics of key proteins (γ-H2AX, 53-BP1, ATM kinase, MDC1) after radiation exposure.
To validate the use of an automated capillary irradiator for consistent DNA double-strand break induction in fingerstick blood samples.
To demonstrate the scalability of the laboratory-based RABiT system for population studies using γ-H2AX as a biomarker.

Main Methods:

Automated sampling of lymphocytes from a RABiT incubator for time-dependent kinetic studies.
Irradiation of fingerstick blood samples using an automated capillary irradiator to induce DNA double-strand breaks.
Quantification of DNA repair proteins (γ-H2AX, 53-BP1, ATM kinase, MDC1) at multiple time points (0.5, 2, 4, 7, 24 h) post-irradiation with 4 Gy γ rays.

Main Results:

Characterization of the time-dependent kinetics of DNA repair proteins γ-H2AX, 53-BP1, ATM kinase, and MDC1 following gamma irradiation.
Successful implementation of an automated system for consistent DNA double-strand break induction in small blood volumes.
Initiation of a population study using γ-H2AX as a biomarker to demonstrate RABiT system scalability.

Conclusions:

The RABiT system effectively measures DNA repair kinetics in lymphocytes, providing valuable data for radiation biodosimetry.
Automated irradiation and sampling techniques ensure consistency and high throughput for laboratory-based studies.
The laboratory-based RABiT system is scalable for population studies, with γ-H2AX serving as a reliable biomarker.