Clustered Regularly Interspaced Short Palindromic Repeats (CRISPR)/CRISPR-associated protein (Cas) is used by some bacteria and most archaea to protect against viral phage intrusion and has recently been adapted to allow for efficient editing of the mammalian genome. Whilst CRISPR/Cas-based technology has been used to modify genes in mammalian cells in vitro, delivery of CRISPR/Cas system into mammalian tissue and/or organs is more difficult and often requires additional vectors. With the use of adeno-associated virus (AAV) gene delivery system, active CRISPR/Cas enzyme can be maintained for an extended period of time and enable efficient editing of genome in the retina in vivo. Herein we outline the method to edit the genome in mouse retina using a dual AAV vector-mediated CRISPR/Cas9 system.
History
Publication title
Retinal Gene Therapy: Methods and Protocols
Editors
CJF Boon, J Wijnholds
Pagination
113-133
ISBN
978-1-4939-7521-1
Department/School
Menzies Institute for Medical Research
Publisher
Humana Press
Place of publication
United States
Extent
27
Rights statement
Copyright 2018 Springer Science+Business Media LLC