A rationally engineered cytosine base editor retains high on-target activity while reducing both DNA and RNA off-target effects Erwei Zuo 1,2,10 ✉, Yidi Sun 3,4,10, Tanglong Yuan 1,10, Bingbing He 2,10, Changyang Zhou 2,10, Wenqin Ying 2, Jing Liu 1, Wu Wei 4,5, Rong Zeng3,6, Yixue Li 4,6,7,8 ✉ and Hui Yang 2,9 &# 9993;
Cytosine base editors (CBEs) offer a powerful tool for correcting point mutations, yet their DNA and RNA off-target activities have caused concerns in biomedical applications. We describe screens of 23 rationally engineered CBE variants, which reveal mutation residues in the predicted DNA-binding site can dra- matically decrease the Cas9-independent off-target effects. Furthermore, we obtained a CBE variant—YE1-BE3-FNLS— that retains high on-target editing efficiency while causing extremely low off-target edits and bystander edits.
去年杨辉发表的 Sicence:
Cytosine base editor generates substantial off-target single-nucleotide variants in mouse embryos Erwei Zuo1,2,*, Yidi Sun3,*, Wu Wei4,5,6,*, Tanglong Yuan2,*, Wenqin Ying1, Hao Sun7, Liyun Yuan4, Lars M. Steinmetz5,8,9,†, Yixue Li4,10,11,&# 8224;, Hui Yang1,†
Abstract Genome editing holds promise for correcting pathogenic mutations. However, it is difficult to determine off-target effects of editing due to single- nucleotide polymorphism in individuals. Here we developed a method named GOTI (genome-wide off-target analysis by two-cell embryo injection) to detect off-target mutations by editing one blastomere of two-cell mouse embryos using either CRISPR-Cas9 or base editors. Comparison of the whole- genome sequences of progeny cells of edited and nonedited blastomeres at embryonic day 14.5 showed that off-target single-nucleotide variants (SNVs) were rare in embryos edited by CRISPR-Cas9 or adenine base editor, with a frequency close to the spontaneous mutation rate. By contrast, cytosine base editing induced SNVs at more than 20-fold higher frequencies, requiring a solution to address its fidelity.
不是内行，看了一些文章的简介，说实话，没看出有什么问题,science文的设计也很精巧 【 在 crispr2016 () 的大作中提到: 】 : 现在杨辉发表的 : Nature Methods: : A rationally engineered cytosine base editor retains high on-target activity : while reducing both DNA and RNA off-target effects : Erwei Zuo 1,2,10 ✉, Yidi Sun 3,4,10, Tanglong Yuan 1,10, : Bingbing He 2,10, Changyang Zhou 2,10, Wenqin Ying 2, Jing Liu 1, Wu : Wei 4,5, Rong Zeng3,6, Yixue Li 4,6,7,8 ✉ and Hui Yang 2,9 &#: 9993; : Cytosine base editors (CBEs) offer a powerful tool for correcting point : mutations, yet their DNA and RNA off-target activities have caused concerns : ...................
现在杨辉发表的
Nature Methods:
A rationally engineered cytosine base editor retains high on-target activity while reducing both DNA and RNA off-target effects
Erwei Zuo 1,2,10 ✉, Yidi Sun 3,4,10, Tanglong Yuan 1,10,
Bingbing He 2,10, Changyang Zhou 2,10, Wenqin Ying 2, Jing Liu 1, Wu Wei 4,5, Rong Zeng3,6, Yixue Li 4,6,7,8 ✉ and Hui Yang 2,9 &#
9993;
Cytosine base editors (CBEs) offer a powerful tool for correcting point
mutations, yet their DNA and RNA off-target activities have caused concerns in biomedical applications. We describe screens of 23 rationally engineered CBE variants, which reveal mutation residues in the predicted DNA-binding
site can dra- matically decrease the Cas9-independent off-target effects.
Furthermore, we obtained a CBE variant—YE1-BE3-FNLS— that retains high on-target editing efficiency while causing extremely low off-target edits and
bystander edits.
去年杨辉发表的
Sicence:
Cytosine base editor generates substantial off-target single-nucleotide
variants in mouse embryos
Erwei Zuo1,2,*, Yidi Sun3,*, Wu Wei4,5,6,*, Tanglong Yuan2,*, Wenqin Ying1, Hao Sun7, Liyun Yuan4, Lars M. Steinmetz5,8,9,†, Yixue Li4,10,11,&#
8224;, Hui Yang1,†
Abstract
Genome editing holds promise for correcting pathogenic mutations. However,
it is difficult to determine off-target effects of editing due to single-
nucleotide polymorphism in individuals. Here we developed a method named
GOTI (genome-wide off-target analysis by two-cell embryo injection) to
detect off-target mutations by editing one blastomere of two-cell mouse
embryos using either CRISPR-Cas9 or base editors. Comparison of the whole-
genome sequences of progeny cells of edited and nonedited blastomeres at
embryonic day 14.5 showed that off-target single-nucleotide variants (SNVs) were rare in embryos edited by CRISPR-Cas9 or adenine base editor, with a
frequency close to the spontaneous mutation rate. By contrast, cytosine base editing induced SNVs at more than 20-fold higher frequencies, requiring a
solution to address its fidelity.
各位怎么看? 杨大师发功往往能够化腐朽为神奇!
不是内行，看了一些文章的简介，说实话，没看出有什么问题,science文的设计也很精巧
【 在 crispr2016 () 的大作中提到: 】
: 现在杨辉发表的
: Nature Methods:
: A rationally engineered cytosine base editor retains high on-target
activity
: while reducing both DNA and RNA off-target effects
: Erwei Zuo 1,2,10 ✉, Yidi Sun 3,4,10, Tanglong Yuan 1,10,
: Bingbing He 2,10, Changyang Zhou 2,10, Wenqin Ying 2, Jing Liu 1, Wu
: Wei 4,5, Rong Zeng3,6, Yixue Li 4,6,7,8 ✉ and Hui Yang 2,9 &#: 9993;
: Cytosine base editors (CBEs) offer a powerful tool for correcting point
: mutations, yet their DNA and RNA off-target activities have caused
concerns
: ...................
给杂志社editor写信或者pubpeer公开质疑吧，在这儿阴阳怪气的显不出本事。
自己有本事造文章就不要怕别人质疑!!!
【 在 finixtree (alix) 的大作中提到: 】
: 给杂志社editor写信或者pubpeer公开质疑吧，在这儿阴阳怪气的显不出本事。