Task Mechanisms of the Repair of HZE Induced DNA Double-Strand Breaks in Human Cells - NNX12AH07G (Completed)
Last Published:  11/23/20 11:55:12 AM (Central)
Short Title: Repair DNA II
Responsible HRP Element: Space Radiation
Collaborating Org(s):
Funding Status: Completed - Task completed and produced a deliverable
Procurement Mechanism(s):
Solicited
Aims:

During our upcoming beam times at the National Space Radiation Laboratory, Brookhaven National Lab, New York, we will test two major hypotheses:

  1. Cells at different phases of cell cycle repair clustered DNA lesions induce HZE particles differently.
  2. The nature of clustered DNA lesions induced by HZE particles is dependent of the physical properties of the particles.

Experiment #1: Establishment of a base damage marker for live cell imaging and evaluation of kinetics of clustered DNA lesions in live cells.

  1. (Identify a suitable base damage marker for live cell imaging.
  2. Evaluate the kinetics of recruitment of different DNA repair factors to the sites of DNA lesions induced by HZE particles in live cells.

Experiment #2: Establish approaches for cell synchronization and study induction and repair of clustered DNA lesions at different phases of cell cycle.

  1. Synchronize cells at different phases of cell cycle.
  2. Identify different phases of cell cycle.
  3. Study induction and repair of clustered DNA lesions at different phases of cell cycle.
  4. Determine cell cycle-dependent recruitment and retention of DNA repair proteins at the sites of clustered DNA lesions.
  5. Determine whether there is a differential repair capability of clustered DNA lesions in G1 vs. S phase of the human HT1080 cells.
  6. Monitor clustered DNA lesions repair in G2 phase of the cell cycle.

Experiment #3: Quantification and imaging analysis of clustered DNA lesions to determine whether the nature of DNA lesions induced by HZE particles is dependent on the physical properties of the HZE particles. These experiments will be carried out by the Co-PI.

  1. Quantify charge and LET dependent induction and repair of clustered DNA lesions.
  2. Determine the nature of clustered DNA lesions induced by different HZE particles.
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