TAILIEUCHUNG - Báo cáo khoa học: Efficient inhibition of b-secretase gene expression in HEK293 cells by tRNAVal-driven and CTE-helicase associated hammerhead ribozymes

Theb-amyloid peptide (Ab) is a major component of toxic amyloid plaques found in the brains of patients with Alzheimer’s liberated by sequential cleavage of amyloid precursor protein (APP) level of Abdepends directly on the hydrolytic activity , b-secretase is an excel-lent target for drug approach based on RNA-cleaving ribozymes was developed to control expression ofb-secretase. | Eur. J. Biochem. 270 3962-3970 2003 FEBS 2003 doi Efficient inhibition of p-secretase gene expression in HEK293 cells by tRNAVal-driven and CTE-helicase associated hammerhead ribozymes Barbara Nawrot1 Slawomir Antoszczvk1. Maria Maszewska1 Tomoko Kuwabara2. Masaki Warashina2 Kazunari Taira2 and Wojciech J. Stec1 1Centre of Molecular and Macromolecular Studies Polish Academy of Sciences Department of Bioorganic Chemistry Lodz Poland 2Gene Function Research Laboratory National Institute of Advanced Industrial Science and Technology AIST Tsukuba Science City Japan The P-amyloid peptide AP is a major component of toxic amyloid plaques found in the brains of patients with Alzheimer s disease. AP is liberated by sequential cleavage of amyloid precursor protein APP by P- and y-secre-tases. The Ivvel of Ap depends directly on the hydrolytic activity of P-secretase. Tliei fl ore. P-secretase is an excellent target for drug design. An ppro ich baved on RNA-cleaving ribozymes was developed to control expression of P-secretase. Two sites ff mRNA coding P-site APP cleaving enzyme were chosen as target sequences for endogenously delivered ribozymes. The rioo yme hasvstie was designed to constitute a catalytic hammerhead core and substrate recognition arms flanked at the 5 -teemincs by tRNAVal and at the 3 -teeminus by constitutive transport element sequences. Rioo _vme cavttvses were cloned into a pUC19 plasmid and used for transient transfection of HEK293 cells. We dymoattraie dial cudi ribozymes efficiently inhibit P-secretase gene expression at both the mRNA up to 95 and the protein up to 90 levels. Iiihíbmon ff P-site APP cleaving enzyme activity directly influences the intra- and extracellular population of AP peptide. TC i leỉ oiv such ribozymes may be considered as molecular tools for silencing the P-secretase activity and further as therapeutic agents for anti-amyloid treatment. Keywords Alzheimer s disease hammerhead ribozyme .

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