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Bacterial Artificial Chromosomes: A Functional Genomics Tool for the Study of Positive-strand RNA Viruses
Published on: December 29, 2015
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The infectious BAC genomic DNA expression library: a high capacity vector system for functional genomics
Michele M P Lufino1, Pauline A H Edser1, Michael A Quail2
1Department of Physiology, Anatomy and Genetics, University of Oxford, Le Gros Clark Building, South Parks Road, Oxford, OX1 3QX, UK.
Scientific Reports
|June 30, 2016
Summary
Researchers developed a novel infectious BAC (iBAC) library using herpes simplex virus-1 amplicon technology. This tool delivers genomic DNA for physiologically relevant gene expression, aiding genetic rescue studies in various cell types.
Area of Science:
- * Molecular Biology
- * Genomics
- * Virology
Background:
- * Gene dosage significantly impacts cellular phenotypes.
- * Traditional cDNA expression systems often exceed physiological protein levels.
- * Genomic DNA expression vectors offer physiologically relevant gene expression by including regulatory elements.
Purpose of the Study:
- * To create the first genomic DNA expression library using herpes simplex virus-1 amplicon technology.
- * To enable the delivery of bacterial artificial chromosomes (BACs) into cells via viral transduction.
- * To provide a new tool for genetic analysis and rescue studies.
Main Methods:
- * Generation of a high-capacity herpes simplex virus-1 amplicon-based library.
- * Packaging of bacterial artificial chromosomes (BACs) into infectious viral particles (iBACs).
- * Delivery of iBACs into Chinese hamster ovary (CHO) and mouse embryonic stem (ES) cells via viral transduction.
Main Results:
- * Creation of an iBAC library with 184,320 clones and an average insert size of 134.5 kb.
- * Successful genetic rescue studies in CHO and ES cells.
- * Demonstrated physiological restoration of Ldlr deficiency and viral receptor expression.
Conclusions:
- * The iBAC library is a pioneering tool for genomic DNA expression.
- * It enables physiologically relevant gene expression and genetic rescue.
- * This resource is now openly available to the research community for advanced genetic analysis.
