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1
Evaluation and Minimization of Cas9-Independent Off-Target DNA Editing by Cytosine Base Editors
In: Nat Biotechnol (2020)
Abstract: Cytosine base editors (CBEs) enable targeted C•G-to-T•A conversions in genomic DNA. Recent studies report that BE3, the original CBE, induces a low frequency of genome-wide Cas9-independent off-target C•G-to-T•A deamination in mouse embryos and in rice. Here we develop multiple rapid, cost-effective methods to screen the propensity of different CBEs to induce Cas9-independent deamination in E. coli and in human cells. We use these assays to identify CBEs with reduced Cas9-independent deamination and validate via whole-genome sequencing that YE1, a narrowed-window CBE variant, displays background levels of Cas9-independent off-target editing. We engineered YE1 variants that retain the substrate-targeting scope of high-activity CBEs while maintaining minimal Cas9-independent off-target editing. The suite of CBEs characterized and engineered in this study collectively offer ~10- to 100-fold lower average Cas9-independent off-target DNA editing while maintaining robust on-target editing at most positions targetable by canonical CBEs, and thus are especially promising for applications in which off-target editing must be minimized.
Keyword: Article
URL: http://www.ncbi.nlm.nih.gov/pubmed/32042165
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC7335424/
https://doi.org/10.1038/s41587-020-0414-6
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2
Neural correlates of children's theory of mind development
In: Child development. - Malden, Ma. [u.a.] : Blackwell 80 (2009) 2, 318-326
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3
Neural correlates of belief- and desire-reasoning
In: Child development. - Malden, Ma. [u.a.] : Blackwell 80 (2009) 4, 1163-1171
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4
Theory of mind development in Chinese children: a meta-analysis of false-belief understanding across cultures and languages
In: Developmental psychology. - Richmond, Va. [u.a.] : American Psychological Association 44 (2008) 2, 523-531
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5
Preschoolers' understanding of knowing-that and knowing-how in the United States and Hong Kong
In: Developmental psychology. - Richmond, Va. [u.a.] : American Psychological Association 41 (2005) 3, 562-573
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6
Preschoolers' understanding of knowing-that and knowing-how in the United States and Hong Kong
In: Developmental psychology. - Richmond, Va. [u.a.] : American Psychological Association 41 (2005) 3, 562-573
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7
Steps in theory-of-mind development for children with deafness or autism
In: Child development. - Malden, Ma. [u.a.] : Blackwell 76 (2005) 2, 502-517
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8
Scaling of theory-of-mind tasks
In: Child development. - Malden, Ma. [u.a.] : Blackwell 75 (2004) 2, 523-541
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9
Automated Suggestions for Miscollocations
In: http://www.cs.rochester.edu/u/tetreaul/bea4/Liu-BEA4.pdf
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10
Automated Suggestions for Miscollocations
In: http://aclweb.org/anthology-new/W/W09/W09-2107.pdf
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