title: Design and cloning of short hairpin RNAs (shRNAs) into a lentiviral silencing vector to study the function of selected proteins in neuronal apoptosis.
abstract: Double-stranded RNA -mediated interference (RNAi ) is a new simple and fast research tool for shutting down genes and characterizes function of their respective proteins. Many strategies for design and delivery of siRNA to target cells are available. Here, we describe the use of lentiviral short hairpin RNA (shRNA) RNA silencing to identify the involvement of d-serine racemase (SR )- an enzyme that syntheses d-serine to modulate glutamate- N-methyl-D-aspartate receptor- in regulating rat cerebellar granule neurons (CGN ) apoptosis. Apoptosis is induced by serum and KCl withdrawal and is detected with fluorometric caspase 3 assay.
authors.affiliation: Department of System Medicine, University of "Tor Vergata" Rome, Via Montpellier 1, Rome, 00137, Italy, cnanda00@uniroma2.it.
authors.forename: Nadia
authors.initials: N
authors.lastname: Canu
mesh_terms: D000818:Animals; D017209:Apoptosis / Q000235:genetics*; D006801:Humans; D016086:Lentivirus / Q000235:genetics; D008967:Molecular Biology / Q000379:methods; D009474:Neurons / Q000378:metabolism*; D034741:RNA, Small Interfering / Q000235:genetics*; D051381:Rats; D016194:Receptors, N-Methyl-D-Aspartate / Q000235:genetics*
chemical_list: D034741:RNA, Small Interfering; D016194:Receptors, N-Methyl-D-Aspartate