1: Nucleic Acids Res. 2001 May 1;29(9):1982-8. 

Condensation by DNA looping facilitates transfer of large DNA molecules into
mammalian cells.

Montigny WJ, Houchens CR, Illenye S, Gilbert J, Coonrod E, Chang YC, Heintz NH.

Department of Pathology, University of Vermont, Burlington, VT 05405, USA.

Experimental studies of complete mammalian genes and other genetic domains are
impeded by the difficulty of introducing large DNA molecules into cells in
culture. Previously we have shown that GST-Z2, a protein that contains three
zinc fingers and a proline-rich multimerization domain from the polydactyl zinc
finger protein RIP60 fused to glutathione S-transferase (GST), mediates DNA
binding and looping in vitro. Atomic force microscopy showed that GST-Z2 is able
to condense 130-150 kb bacterial artificial chromosomes (BACs) into protein-DNA
complexes containing multiple DNA loops. Condensation of the DNA loops onto the
Z2 protein-BAC DNA core complexes with cationic lipid resulted in particles that
were readily transferred into multiple cell types in culture. Transfer of total
genomic linear DNA containing amplified DHFR genes into DHFR(-) cells by GST-Z2
resulted in a 10-fold higher transformation rate than calcium phosphate
co-precipitation. Chinese hamster ovarian cells transfected with a BAC
containing the human TP53 gene locus expressed p53, showing native promoter
elements are active after GST-Z2-mediated gene transfer. Because DNA
condensation by GST-Z2 does not require the introduction of specific recognition
sequences into the DNA substrate, condensation by the Z2 domain of RIP60 may be
used in conjunction with a variety of other agents to provide a flexible and
efficient non-viral platform for the delivery of large genes into mammalian
cells.

PMID: 11328883 [PubMed - indexed for MEDLINE]


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