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High Capacity Extrachromosomal Gene Expression Vectors

  • Olivia HibbittEmail author
  • Richard Wade-Martins
Protocol
Part of the Methods in Molecular Biology book series (MIMB, volume 738)

Abstract

Extrachromosomal gene expression vectors that contain native genomic gene expression elements have numerous advantages over traditional integrating mini-gene vectors. In this protocol chapter we describe our work using episomal vectors where expression of a cDNA is controlled by a 10 kB piece of genomic DNA encompassing the promoter of the low density lipoprotein receptor. We explain methods to sub-clone large genomic inserts into gene expression vectors. We also illustrate various methods employed to ascertain whether expression from these vectors is robust and physiologically relevant by investigating their sensitivity to changes in cellular milieu. Delivery of gene expression vectors in vivo is also described using hydrodynamic tail vein injection, a high pressure, high volume tail vein injection used for liver-directed gene transfer.

Key words

Episomal Hydrodynamic tail vein injection LDLR Familial hypercholesterolaemia Live imaging Cholesterol Genomic promoter Luciferase 

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Copyright information

© Springer Science+Business Media, LLC 2011

Authors and Affiliations

  1. 1.Department of Anatomy and GeneticsUniversity of OxfordOxfordUK

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