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Salt controls size of DNA structures, could improve gene therapy

al parameter in studying molecules in solution, particularly such highly charged molecules as DNA."

Protocols for preparing DNA for delivery to cells often call for salt conditions that differ from those DNA encounters when injected into body tissues, Hud noted. "If you change the salt conditions during DNA delivery, it will change particle size and have a dramatic effect on the efficiency of gene delivery," he added. "This could explain why some researchers aren't getting as good a rate of transfection (the incorporation of DNA into a cell) as they should."

In the study reported in PNAS, Hud and his Ph.D. students Christine Conwell and Igor Vilfan also describe using the positively charged, inorganic molecule hexammine cobalt (III) to condense a DNA molecule containing a specially designed sequence. The synthetic sequence causes a region of the DNA molecule to bend into two loops of 25 nanometers each in diameter. In other words, these nanoscale loops were "programmed" into the DNA sequence.

Hud theorized years ago that the spontaneous formation of loops along DNA is the first step necessary for toroid formation, and a key factor in determining toroid size, he said. "Now, we've made these loops always present there," Hud explained. "When we add positively charged molecules that bind to the DNA, the loops provide a built-in starting point for DNA condensation. The loops also act as a template upon which the rest of the DNA rolls up to form a toroid. The toroid forms because the positively charged molecules make DNA want to stick to itself. and we found that our static loops reduce DNA particle size and tighten particle distribution."

Hud describes as serendipitous the additional finding that lowering the salt concentration in DNA solution also reduces the size of toroids. Together, these results helped Hud's team develop models for DNA toroid formation. The researchers' data can now serve as a test for theoretical models of DNA condensation, t
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Contact: Jane Sanders
jane.sanders@edi.gatech.edu
404-894-2214
Georgia Institute of Technology Research News
29-Jul-2003


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