title: A 5' Noncoding Exon Containing Engineered Intron Enhances Transgene Expression from Recombinant AAV Vectors in vivo.
abstract: We previously developed a mini-intronic plasmid (MIP) expression system in which the essential bacterial elements for plasmid replication and selection are placed within an engineered intron contained within a universal 5' UTR noncoding exon. Like minicircle DNA plasmids (devoid of bacterial backbone sequences), MIP plasmids overcome transcriptional silencing of the transgene. However, in addition MIP plasmids increase transgene expression by 2 and often >10 times higher than minicircle vectors in vivo and in vitro. Based on these findings, we examined the effects of the MIP intronic sequences in a recombinant adeno-associated virus (AAV) vector system. Recombinant AAV vectors containing an intron with a bacterial replication origin and bacterial selectable marker increased transgene expression by 40 to 100 times in vivo when compared with conventional AAV vectors. Therefore, inclusion of this noncoding exon/intron sequence upstream of the coding region can substantially enhance AAV-mediated gene expression in vivo.
authors.affiliation: 1 Department of Pediatrics, Stanford University , Palo Alto, California.|2 Department of Genetics, Stanford University , Palo Alto, California.
authors.forename: Jiamiao
authors.initials: J
authors.lastname: Lu
authors.affiliation: 3 Nature Technology Corporation , Lincoln, Nebraska.
authors.forename: James A
authors.initials: JA
authors.lastname: Williams
authors.affiliation: 3 Nature Technology Corporation , Lincoln, Nebraska.
authors.forename: Jeremy
authors.initials: J
authors.lastname: Luke
authors.affiliation: 1 Department of Pediatrics, Stanford University , Palo Alto, California.|2 Department of Genetics, Stanford University , Palo Alto, California.
authors.forename: Feijie
authors.initials: F
authors.lastname: Zhang
authors.affiliation: 1 Department of Pediatrics, Stanford University , Palo Alto, California.|2 Department of Genetics, Stanford University , Palo Alto, California.
authors.forename: Kirk
authors.initials: K
authors.lastname: Chu
authors.affiliation: 1 Department of Pediatrics, Stanford University , Palo Alto, California.|2 Department of Genetics, Stanford University , Palo Alto, California.
authors.forename: Mark A
authors.initials: MA
authors.lastname: Kay
mesh_terms: D020121:5' Untranslated Regions / Q000235:genetics*; D000818:Animals; D004261:DNA Replication; D000229:Dependovirus / Q000235:genetics*; D005091:Exons / Q000235:genetics*; D005260:Female; D017930:Genes, Reporter / Q000502:physiology*; D005822:Genetic Vectors / Q000008:administration & dosage / Q000235:genetics*; D006801:Humans; D007438:Introns / Q000235:genetics*; D051379:Mice; D008810:Mice, Inbred C57BL; D011995:Recombination, Genetic; D019076:Transgenes / Q000502:physiology*
chemical_list: D020121:5' Untranslated Regions
keywords: AAV vectors; enhance transgene expression; intron; mini-intronic plasmid; miniorigins