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Journal of Cell Science, Vol 112, Issue 12 2033-2041, Copyright © 1999 by Company of Biologists

JOURNAL ARTICLES

A nuclear localization signal can enhance both the nuclear transport and expression of 1 kb DNA

JJ Ludtke, G Zhang, MG Sebestyen and JA Wolff
Departments of Pediatrics and Medical Genetics, Waisman Center, University of Wisconsin-Madison, Madison, WI 53705, USA.

Although the entry of DNA into the nucleus is a crucial step of non-viral gene delivery, fundamental features of this transport process have remained unexplored. This study analyzed the effect of linear double stranded DNA size on its passive diffusion, its active transport and its NLS-assisted transport. The size limit for passive diffusion was found to be between 200 and 310 bp. DNA of 310-1500 bp entered the nuclei of digitonin treated cells in the absence of cytosolic extract by an active transport process. Both the size limit and the intensity of DNA nuclear transport could be increased by the attachment of strong nuclear localization signals. Conjugation of a 900 bp expression cassette to nuclear localization signals increased both its nuclear entry and expression in microinjected, living cells.
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