Cookies on this website

We use cookies to ensure that we give you the best experience on our website. If you click 'Accept all cookies' we'll assume that you are happy to receive all cookies and you won't see this message again. If you click 'Reject all non-essential cookies' only necessary cookies providing core functionality such as security, network management, and accessibility will be enabled. Click 'Find out more' for information on how to change your cookie settings.

As gene therapy of choroideremia is becoming a clinical reality, there is a need for reliable and sensitive assays to determine the expression of exogenously delivered Rab Escort Protein-1 (REP1), in particular to test new gene therapy vectors and as a quality control screen for clinical vector stocks. Here we describe an in vitro protocol to test transgene expression following AAV2/2-REP1 transduction of a human cell line. Gene augmentation can be confirmed by western blot and quantification of the fold-increase of human REP1 levels over untransduced controls.

Original publication

DOI

10.1007/978-1-4939-7522-8_7

Type

Chapter

Publication Date

2018

Volume

1715

Pages

89 - 97

Keywords

AAV, Choroideremia, Gene augmentation, Gene therapy, Human REP1, Western blot, Adaptor Proteins, Signal Transducing, Choroideremia, Dependovirus, Gene Transfer Techniques, Genetic Therapy, Genetic Vectors, HEK293 Cells, Humans, Retina