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优化的碱基编辑器可实现细胞、类器官和小鼠中的高效编辑。

Optimized base editors enable efficient editing in cells, organoids and mice.

机构信息

Sandra and Edward Meyer Cancer Center, Department of Medicine, Weill Cornell Medicine, New York, New York, USA.

Weill Cornell/Rockefeller/Sloan-Kettering Tri-Institutional MD-PhD program, New York, New York, USA.

出版信息

Nat Biotechnol. 2018 Oct;36(9):888-893. doi: 10.1038/nbt.4194. Epub 2018 Jul 3.


DOI:10.1038/nbt.4194
PMID:29969439
原文链接:https://pmc.ncbi.nlm.nih.gov/articles/PMC6130889/
Abstract

CRISPR base editing enables the creation of targeted single-base conversions without generating double-stranded breaks. However, the efficiency of current base editors is very low in many cell types. We reengineered the sequences of BE3, BE4Gam, and xBE3 by codon optimization and incorporation of additional nuclear-localization sequences. Our collection of optimized constitutive and inducible base-editing vector systems dramatically improves the efficiency by which single-nucleotide variants can be created. The reengineered base editors enable target modification in a wide range of mouse and human cell lines, and intestinal organoids. We also show that the optimized base editors mediate efficient in vivo somatic editing in the liver in adult mice.

摘要

CRISPR 碱基编辑可在不产生双链断裂的情况下实现靶向单碱基转换。然而,目前的碱基编辑器在许多细胞类型中的效率非常低。我们通过密码子优化和额外核定位序列的掺入,重新设计了 BE3、BE4Gam 和 xBE3 的序列。我们优化的组成型和诱导型碱基编辑载体系统显著提高了创建单核苷酸变体的效率。经重新设计的碱基编辑器可在广泛的小鼠和人类细胞系以及肠道类器官中实现靶标修饰。我们还表明,优化后的碱基编辑器可介导成年小鼠肝脏中的高效体内体细胞编辑。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/a9a0/6130889/c2bcd84cc88f/nihms-977467-f0003.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/a9a0/6130889/58f7b08ae6d3/nihms-977467-f0001.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/a9a0/6130889/c8dd2ab4fc9b/nihms-977467-f0002.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/a9a0/6130889/c2bcd84cc88f/nihms-977467-f0003.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/a9a0/6130889/58f7b08ae6d3/nihms-977467-f0001.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/a9a0/6130889/c8dd2ab4fc9b/nihms-977467-f0002.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/a9a0/6130889/c2bcd84cc88f/nihms-977467-f0003.jpg

相似文献

[1]
Optimized base editors enable efficient editing in cells, organoids and mice.

Nat Biotechnol. 2018-7-3

[2]
[CRISPR/Cas-mediated DNA base editing technology and its application in biomedicine and agriculture].

Sheng Wu Gong Cheng Xue Bao. 2021-9-25

[3]
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Biochem Biophys Res Commun. 2024-12-20

[4]
Improving cytidine and adenine base editors by expression optimization and ancestral reconstruction.

Nat Biotechnol. 2018-5-29

[5]
Rapid Vector Construction and Assessment of BE3 and Target-AID C to T Base Editing Systems in Rice Protoplasts.

Methods Mol Biol. 2021

[6]
Improving the Precision of Base Editing by Bubble Hairpin Single Guide RNA.

mBio. 2021-4-20

[7]
Efficient Generation and Correction of Mutations in Human iPS Cells Utilizing mRNAs of CRISPR Base Editors and Prime Editors.

Genes (Basel). 2020-5-6

[8]
In vivo somatic cell base editing and prime editing.

Mol Ther. 2021-11-3

[9]
In Vivo Base Editing of PCSK9 (Proprotein Convertase Subtilisin/Kexin Type 9) as a Therapeutic Alternative to Genome Editing.

Arterioscler Thromb Vasc Biol. 2017-9

[10]
Off-Target Editing by CRISPR-Guided DNA Base Editors.

Biochemistry. 2019-8-26

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[2]
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[3]
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[4]
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[5]
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[6]
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[7]
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[8]
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Cell Chem Biol. 2025-6-19

[9]
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[10]
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本文引用的文献

[1]
Improving cytidine and adenine base editors by expression optimization and ancestral reconstruction.

Nat Biotechnol. 2018-5-29

[2]
Adenine base editing in mouse embryos and an adult mouse model of Duchenne muscular dystrophy.

Nat Biotechnol. 2018-4-27

[3]
Evolved Cas9 variants with broad PAM compatibility and high DNA specificity.

Nature. 2018-2-28

[4]
Programmable base editing of A•T to G•C in genomic DNA without DNA cleavage.

Nature. 2017-11-23

[5]
Rescue of high-specificity Cas9 variants using sgRNAs with matched 5' nucleotides.

Genome Biol. 2017-11-15

[6]
Structure-guided chemical modification of guide RNA enables potent non-viral in vivo genome editing.

Nat Biotechnol. 2017-12

[7]
RNA editing with CRISPR-Cas13.

Science. 2017-11-24

[8]
Improved base excision repair inhibition and bacteriophage Mu Gam protein yields C:G-to-T:A base editors with higher efficiency and product purity.

Sci Adv. 2017-8-30

[9]
R-Spondin chromosome rearrangements drive Wnt-dependent tumour initiation and maintenance in the intestine.

Nat Commun. 2017-7-11

[10]
mTOR signaling mediates resistance to tankyrase inhibitors in Wnt-driven colorectal cancer.

Oncotarget. 2017-7-18

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