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Methods for In Vivo CRISPR/Cas Editing of the Adult Murine Retina

Hung, SS, Li, F ORCID: 0000-0001-7273-7317, Wang, J-H, King, AE ORCID: 0000-0003-1792-0965, Bui, BV and Liu, G ORCID: 0000-0003-3379-724X 2018 , 'Methods for In Vivo CRISPR/Cas Editing of the Adult Murine Retina', in CJF Boon and J Wijnholds (eds.), Retinal Gene Therapy: Methods and Protocols , Humana Press, United States, pp. 113-133.

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Abstract

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.

Item Type: Book Section
Authors/Creators:Hung, SS and Li, F and Wang, J-H and King, AE and Bui, BV and Liu, G
Keywords: CRISPR/Cas9, genome editing, gene delivery, AAV, retina
Publisher: Humana Press
DOI / ID Number: 10.1007/978-1-4939-7522-8_9
Copyright Information:

Copyright 2018 Springer Science+Business Media LLC

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