Enhanced Genome Editing Tools For Multi‐Gene Deletion Knock‐Out Approaches Using Paired CRISPR sgRNAs in CHO Cells

Enhanced Genome Editing Tools For Multi‐Gene Deletion Knock‐Out Approaches Using Paired... IntroductionChinese Hamster Ovary (CHO) cells are the most widely used expression system for the production of complex therapeutic proteins since their establishment as cell factories in 1987. Despite the importance of CHO cells for industry, it was only recently that genomic and transcriptomic data of high quality was acquired. However, now, that such data sets are available, the focus of optimization of cell performance has shifted from the original approach of optimizing media, cell selection procedures, and production process conditions, toward directed cell engineering.In parallel, the development of tools for targeted cell engineering has made a tremendous leap forward with the introduction of nucleases derived from bacterial clustered regularly interspaced short palindromic repeat (CRISPR) immune systems. Cas9, originally identified as the effector endonuclease of Streptococcus pyogenes, has been proven to be an efficient editing tool in many mammalian cells using a single chimeric RNA guide (single guide RNA − sgRNA). Cas9 introduces DNA double strand breaks (DSB), which are repaired by either homology directed repair or non‐homologous end‐joining (NHEJ), with the latter being the more frequent mechanism in mammalian cells. NHEJ usually results in the insertion or deletion (InDel) of nucleotides at the targeted site. With CRISPR/Cas9, genes can http://www.deepdyve.com/assets/images/DeepDyve-Logo-lg.png Biotechnology Journal Wiley

Enhanced Genome Editing Tools For Multi‐Gene Deletion Knock‐Out Approaches Using Paired CRISPR sgRNAs in CHO Cells

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Publisher
Wiley Subscription Services, Inc., A Wiley Company
Copyright
© 2018 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim
ISSN
1860-6768
eISSN
1860-7314
D.O.I.
10.1002/biot.201700211
Publisher site
See Article on Publisher Site

Abstract

IntroductionChinese Hamster Ovary (CHO) cells are the most widely used expression system for the production of complex therapeutic proteins since their establishment as cell factories in 1987. Despite the importance of CHO cells for industry, it was only recently that genomic and transcriptomic data of high quality was acquired. However, now, that such data sets are available, the focus of optimization of cell performance has shifted from the original approach of optimizing media, cell selection procedures, and production process conditions, toward directed cell engineering.In parallel, the development of tools for targeted cell engineering has made a tremendous leap forward with the introduction of nucleases derived from bacterial clustered regularly interspaced short palindromic repeat (CRISPR) immune systems. Cas9, originally identified as the effector endonuclease of Streptococcus pyogenes, has been proven to be an efficient editing tool in many mammalian cells using a single chimeric RNA guide (single guide RNA − sgRNA). Cas9 introduces DNA double strand breaks (DSB), which are repaired by either homology directed repair or non‐homologous end‐joining (NHEJ), with the latter being the more frequent mechanism in mammalian cells. NHEJ usually results in the insertion or deletion (InDel) of nucleotides at the targeted site. With CRISPR/Cas9, genes can

Journal

Biotechnology JournalWiley

Published: Jan 1, 2018

Keywords: ; ; ;

References

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