title: The infectious BAC genomic DNA expression library: a high capacity vector system for functional genomics.
abstract: Gene dosage plays a critical role in a range of cellular phenotypes, yet most cellular expression systems use heterologous cDNA-based vectors which express proteins well above physiological levels. In contrast, genomic DNA expression vectors generate physiologically-relevant levels of gene expression by carrying the whole genomic DNA locus of a gene including its regulatory elements. Here we describe the first genomic DNA expression library generated using the high-capacity herpes simplex virus-1 amplicon technology to deliver bacterial artificial chromosomes (BACs) into cells by viral transduction. The infectious BAC (iBAC) library contains 184,320 clones with an average insert size of 134.5 kb. We show in a Chinese hamster ovary (CHO) disease model cell line and mouse embryonic stem (ES) cells that this library can be used for genetic rescue studies in a range of contexts including the physiological restoration of Ldlr deficiency, and viral receptor expression. The iBAC library represents an important new genetic analysis tool openly available to the research community.
authors.affiliation: Department of Physiology, Anatomy and Genetics, University of Oxford, Le Gros Clark Building, South Parks Road, Oxford, OX1 3QX, UK.
authors.forename: Michele M P
authors.initials: MM
authors.lastname: Lufino
authors.affiliation: Department of Physiology, Anatomy and Genetics, University of Oxford, Le Gros Clark Building, South Parks Road, Oxford, OX1 3QX, UK.
authors.forename: Pauline A H
authors.initials: PA
authors.lastname: Edser
authors.affiliation: Wellcome Trust Sanger Institute, Hinxton, Cambridgeshire, CB10 1SA, UK.
authors.forename: Michael A
authors.initials: MA
authors.lastname: Quail
authors.affiliation: Wellcome Trust Sanger Institute, Hinxton, Cambridgeshire, CB10 1SA, UK.
authors.forename: Stephen
authors.initials: S
authors.lastname: Rice
authors.affiliation: Wellcome Trust Sanger Institute, Hinxton, Cambridgeshire, CB10 1SA, UK.
authors.forename: David J
authors.initials: DJ
authors.lastname: Adams
authors.affiliation: Department of Physiology, Anatomy and Genetics, University of Oxford, Le Gros Clark Building, South Parks Road, Oxford, OX1 3QX, UK.
authors.forename: Richard
authors.initials: R
authors.lastname: Wade-Martins
mesh_terms: D000818:Animals; D016466:CHO Cells; D022202:Chromosomes, Artificial, Bacterial / Q000235:genetics*; D003412:Cricetulus; D004926:Escherichia coli; D015870:Gene Expression; D015723:Gene Library; D018259:Herpesvirus 1, Human / Q000235:genetics*; D051379:Mice; D000066450:Mouse Embryonic Stem Cells; D014161:Transduction, Genetic / Q000379:methods*