sgBE: a structure-guided design of sgRNA architecture specifies base editing window and enables simultaneous conversion of cytosine and adenosine

We present a base editing system, in which base editors are attached to different sites of sgRNA scaffold (sgBE). Each independent sgBE has its own specific editing pattern for a given target site. Among tested sgBEs, sgBE-SL4, in which deaminase is attached to the last stem-loop of sgRNA, yields th...

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Autores principales: Wang, Yanhong, Zhou, Lifang, Tao, Rui, Liu, Nan, Long, Jie, Qin, Fengming, Tang, Wenling, Yang, Yang, Chen, Qiang, Yao, Shaohua
Formato: Online Artículo Texto
Lenguaje:English
Publicado: BioMed Central 2020
Materias:
Method
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC7453718/
https://www.ncbi.nlm.nih.gov/pubmed/32859244
http://dx.doi.org/10.1186/s13059-020-02137-6
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author Wang, Yanhong
Zhou, Lifang
Tao, Rui
Liu, Nan
Long, Jie
Qin, Fengming
Tang, Wenling
Yang, Yang
Chen, Qiang
Yao, Shaohua
author_facet Wang, Yanhong
Zhou, Lifang
Tao, Rui
Liu, Nan
Long, Jie
Qin, Fengming
Tang, Wenling
Yang, Yang
Chen, Qiang
Yao, Shaohua
author_sort Wang, Yanhong
collection PubMed
description We present a base editing system, in which base editors are attached to different sites of sgRNA scaffold (sgBE). Each independent sgBE has its own specific editing pattern for a given target site. Among tested sgBEs, sgBE-SL4, in which deaminase is attached to the last stem-loop of sgRNA, yields the highest editing efficiency in the window several nucleotides next to the one edited by BE3. sgBE enables the simultaneous editing of adenine and cytosine. Finally, in order to facilitate in vivo base editing, we extend our sgBE system to an AAV-compatible Cas9, SaCas9 (Staphylococcus aureus), and observe robust base editing.
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spelling pubmed-74537182020-08-28 sgBE: a structure-guided design of sgRNA architecture specifies base editing window and enables simultaneous conversion of cytosine and adenosine Wang, Yanhong Zhou, Lifang Tao, Rui Liu, Nan Long, Jie Qin, Fengming Tang, Wenling Yang, Yang Chen, Qiang Yao, Shaohua Genome Biol Method We present a base editing system, in which base editors are attached to different sites of sgRNA scaffold (sgBE). Each independent sgBE has its own specific editing pattern for a given target site. Among tested sgBEs, sgBE-SL4, in which deaminase is attached to the last stem-loop of sgRNA, yields the highest editing efficiency in the window several nucleotides next to the one edited by BE3. sgBE enables the simultaneous editing of adenine and cytosine. Finally, in order to facilitate in vivo base editing, we extend our sgBE system to an AAV-compatible Cas9, SaCas9 (Staphylococcus aureus), and observe robust base editing. BioMed Central 2020-08-28 /pmc/articles/PMC7453718/ /pubmed/32859244 http://dx.doi.org/10.1186/s13059-020-02137-6 Text en © The Author(s) 2020 Open AccessThis article is licensed under a Creative Commons Attribution 4.0 International License, which permits use, sharing, adaptation, distribution and reproduction in any medium or format, as long as you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons licence, and indicate if changes were made. The images or other third party material in this article are included in the article's Creative Commons licence, unless indicated otherwise in a credit line to the material. If material is not included in the article's Creative Commons licence and your intended use is not permitted by statutory regulation or exceeds the permitted use, you will need to obtain permission directly from the copyright holder. To view a copy of this licence, visit http://creativecommons.org/licenses/by/4.0/. The Creative Commons Public Domain Dedication waiver (http://creativecommons.org/publicdomain/zero/1.0/) applies to the data made available in this article, unless otherwise stated in a credit line to the data.
spellingShingle Method
Wang, Yanhong
Zhou, Lifang
Tao, Rui
Liu, Nan
Long, Jie
Qin, Fengming
Tang, Wenling
Yang, Yang
Chen, Qiang
Yao, Shaohua
sgBE: a structure-guided design of sgRNA architecture specifies base editing window and enables simultaneous conversion of cytosine and adenosine
title sgBE: a structure-guided design of sgRNA architecture specifies base editing window and enables simultaneous conversion of cytosine and adenosine
title_full sgBE: a structure-guided design of sgRNA architecture specifies base editing window and enables simultaneous conversion of cytosine and adenosine
title_fullStr sgBE: a structure-guided design of sgRNA architecture specifies base editing window and enables simultaneous conversion of cytosine and adenosine
title_full_unstemmed sgBE: a structure-guided design of sgRNA architecture specifies base editing window and enables simultaneous conversion of cytosine and adenosine
title_short sgBE: a structure-guided design of sgRNA architecture specifies base editing window and enables simultaneous conversion of cytosine and adenosine
title_sort sgbe: a structure-guided design of sgrna architecture specifies base editing window and enables simultaneous conversion of cytosine and adenosine
topic Method
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC7453718/
https://www.ncbi.nlm.nih.gov/pubmed/32859244
http://dx.doi.org/10.1186/s13059-020-02137-6
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