Directed evolution using dCas9-targeted somatic hypermutation in mammalian cells

Recruiting a hyperactive cytidine deaminase via the guide RNA to dCas9 allows for the introduction of diverse point mutations at the CRISPR target locus to create complex libraries of variants for protein engineering or dissection of protein function. Engineering and study of protein function by dir...

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Bibliographic Details
Published in:	Nature methods Vol. 13; no. 12; pp. 1036 - 1042
Main Authors:	Hess, Gaelen T, Frésard, Laure, Han, Kyuho, Lee, Cameron H, Li, Amy, Cimprich, Karlene A, Montgomery, Stephen B, Bassik, Michael C
Format:	Journal Article
Language:	English
Published:	New York Nature Publishing Group US 01.12.2016 Nature Publishing Group
Subjects:	631/208 631/61 Analysis Bioinformatics Biological Microscopy Biological Techniques Biomedical Engineering/Biotechnology Bortezomib - pharmacology Cells Clustered Regularly Interspaced Short Palindromic Repeats - genetics CRISPR-Associated Proteins - genetics Cytidine Deaminase - genetics Deoxyribonucleic acid Directed Molecular Evolution - methods DNA Drug Resistance - genetics Evolutionary biology Gene mutation Genetic variance Green Fluorescent Proteins - genetics Health aspects Humans K562 Cells Levivirus - genetics Life Sciences Mammals Mutation Point Mutation Proteasome Endopeptidase Complex - genetics Protein Engineering - methods Proteins Proteomics RNA, Guide, CRISPR-Cas Systems - genetics Tissue engineering Transcription (Genetics) United States
ISSN:	1548-7091, 1548-7105, 1548-7105
Online Access:	Get full text
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