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A Novel G418 Conjugate Results In Targeted Selection of Genetically Protected Hepatocytes without Bystander Toxicity (CROSBI ID 140552)

Prilog u časopisu | izvorni znanstveni rad | međunarodna recenzija

Volarević, Martina ; Wu, Catherine H. ; Smolić, Robert ; Andorfer, John H. ; Wu, George Y. A Novel G418 Conjugate Results In Targeted Selection of Genetically Protected Hepatocytes without Bystander Toxicity // Bioconjugate chemistry, 18 (2007), 6; 1965-1971. doi: 10.1021/bc700277d

Podaci o odgovornosti

Volarević, Martina ; Wu, Catherine H. ; Smolić, Robert ; Andorfer, John H. ; Wu, George Y.

engleski

A Novel G418 Conjugate Results In Targeted Selection of Genetically Protected Hepatocytes without Bystander Toxicity

G418, an aminoglycoside neomycin analogue, is an antimicrobial agent that interferes with protein synthesis and has been used extensively for selection of mammalian cell lines that possess neomycin resistance (NR). It is potent and nonspecific in its effects that occur through tight binding to ribosomal elements. Because of the potent intracellular effect, we wondered whether G418 could be used to select a specific cell type based on receptor-mediated endocytosis. The objective of this study was to target G418 specifically to liver cells via asialoglycoprotein receptors (AsGR) which are known to be highly selective for these cells. A novel G418 conjugate was synthesized chemically by coupling G418 to a galactose-terminating carrier protein, asialoorosomucoid (AsOR), in a molar ratio of 5:1. AsOR– G418 conjugates inhibited viability of AsGR (+) cells by 84.3%, while inhibition in AsGR (– ) cells was only by 19%. In AsGR (+) cells, stably transfected with a NR gene, the conjugate decreased viability by less than 9%. Furthermore, incubation of conjugate in cocultures of AsGR (+), and AsGR (– ) cells did not result in the loss of viability of neighboring AsGR (– ) cells. Our data demonstrate for the first time that G418 can be covalently bound to AsOR to form a conjugate for hepatocyte-specific targeting and toxicity. AsOR– G418 conjugates may be useful tools for genetic manipulation of human liver cells in the presence of nonhepatic cells.

G418; targeted selection; bystander toxicity; receptor-mediated endocytosis

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Podaci o izdanju

18 (6)

2007.

1965-1971

objavljeno

1043-1802

10.1021/bc700277d

Povezanost rada

Temeljne medicinske znanosti

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