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      Comparison of cytosine base editors and development of the BEable-GPS database for targeting pathogenic SNVs

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          Abstract

          A variety of base editors have been developed to achieve C-to-T editing in different genomic contexts. Here, we compare a panel of five base editors on their C-to-T editing efficiencies and product purity at commonly editable sites, including some human pathogenic C-to-T mutations. We further profile the accessibilities of 20 base editors to all possible pathogenic mutations in silico. Finally, we build the BEable-GPS (Base Editable prediction of Global Pathogenic SNVs) database for users to select proper base editors to model or correct disease-related mutations. The in vivo comparison and in silico profiling catalog the availability of base editors and their broad applications in biomedical studies.

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          Most cited references9

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          CRISPR-Based Technologies for the Manipulation of Eukaryotic Genomes.

          The CRISPR-Cas9 RNA-guided DNA endonuclease has contributed to an explosion of advances in the life sciences that have grown from the ability to edit genomes within living cells. In this Review, we summarize CRISPR-based technologies that enable mammalian genome editing and their various applications. We describe recent developments that extend the generality, DNA specificity, product selectivity, and fundamental capabilities of natural CRISPR systems, and we highlight some of the remarkable advancements in basic research, biotechnology, and therapeutics science that these developments have facilitated.
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            Increasing the genome-targeting scope and precision of base editing with engineered Cas9-cytidine deaminase fusions

            Base editing is a recently developed approach to genome editing that uses a fusion protein containing a catalytically defective Streptococcus pyogenes Cas9, a cytidine deaminase, and an inhibitor of base excision repair to induce programmable, single-nucleotide changes in the DNA of living cells without generating double-strand DNA breaks, without requiring a donor DNA template, and without inducing an excess of stochastic insertions and deletions 1 . Here we report the development of five new C→T (or G→A) base editors that use natural and engineered Cas9 variants with different protospacer-adjacent motif (PAM) specificities to expand the number of sites that can be targeted by base editing by 2.5-fold. Additionally, we engineered new base editors containing mutated cytidine deaminase domains that narrow the width of the apparent editing window from approximately 5 nucleotides to as little as 1 to 2 nucleotides, enabling the discrimination of neighboring C nucleotides that would previously be edited with comparable efficiency, thereby doubling the number of disease-associated target Cs that can be corrected preferentially over nearby non-target Cs. Collectively, these developments substantially increase the targeting scope of base editing and establish the modular nature of base editors.
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              Base editing with a Cpf1–cytidine deaminase fusion

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                Author and article information

                Contributors
                chenjia@shanghaitech.edu.cn
                liyang@picb.ac.cn
                Journal
                Genome Biol
                Genome Biol
                Genome Biology
                BioMed Central (London )
                1474-7596
                1474-760X
                23 October 2019
                23 October 2019
                2019
                : 20
                : 218
                Affiliations
                [1 ]ISNI 0000 0004 1797 8419, GRID grid.410726.6, CAS Key Laboratory of Computational Biology, CAS-MPG Partner Institute for Computational Biology, Shanghai Institute of Nutrition and Health, Shanghai Institutes for Biological Sciences, , University of Chinese Academy of Sciences, Chinese Academy of Sciences, ; Shanghai, 200031 China
                [2 ]GRID grid.440637.2, School of Life Science and Technology, , ShanghaiTech University, ; Shanghai, 201210 China
                [3 ]ISNI 0000000119573309, GRID grid.9227.e, CAS Center for Excellence in Molecular Cell Science, Shanghai Institute of Biochemistry and Cell Biology, , Chinese Academy of Sciences, ; Shanghai, 200031 China
                [4 ]ISNI 0000 0004 1797 8419, GRID grid.410726.6, University of Chinese Academy of Sciences, ; Beijing, 100049 China
                [5 ]GRID grid.440637.2, Shanghai Institute for Advanced Immunochemical Studies, , ShanghaiTech University, ; Shanghai, 201210 China
                Author information
                http://orcid.org/0000-0001-8833-7473
                Article
                1839
                10.1186/s13059-019-1839-4
                6806563
                31647030
                01696d55-b7d5-403b-8147-7a7e3c3e079e
                © The Author(s). 2019

                Open AccessThis article is distributed under the terms of the Creative Commons Attribution 4.0 International License ( http://creativecommons.org/licenses/by/4.0/), which permits unrestricted use, distribution, and reproduction in any medium, provided you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons license, and indicate if changes were made. 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.

                History
                : 29 May 2019
                : 27 September 2019
                Funding
                Funded by: FundRef http://dx.doi.org/10.13039/501100002855, Ministry of Science and Technology of the People's Republic of China;
                Award ID: 2018YFA070227
                Award ID: 2018YFC1004602
                Award Recipient :
                Funded by: FundRef http://dx.doi.org/10.13039/501100001809, National Natural Science Foundation of China;
                Award ID: 31730111
                Award ID: 31822016
                Award ID: 81872305
                Award ID: 31600654
                Award Recipient :
                Categories
                Short Report
                Custom metadata
                © The Author(s) 2019

                Genetics
                base editing,base editor,crispr/cas,cytidine deaminase,pathogenic mutation
                Genetics
                base editing, base editor, crispr/cas, cytidine deaminase, pathogenic mutation

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