AcrIIA5 Suppresses Base Editors and Reduces Their Off-Target Effects
The CRISPR/nCas9-based cytosine base editors (CBEs) and adenine base editors (ABEs) are capable of catalyzing C•G to T•A or A•T to G•C conversions, respectively, and have become new, powerful tools for achieving precise genetic changes in a wide range of organisms. These base editors hold great prom...
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| Format: | Article |
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MDPI AG
2020-07-01
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| Series: | Cells |
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| Online Access: | https://www.mdpi.com/2073-4409/9/8/1786 |
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| author | Mingming Liang Tingting Sui Zhiquan Liu Mao Chen Hongmei Liu Huanhuan Shan Liangxue Lai Zhanjun Li |
| author_facet | Mingming Liang Tingting Sui Zhiquan Liu Mao Chen Hongmei Liu Huanhuan Shan Liangxue Lai Zhanjun Li |
| author_sort | Mingming Liang |
| collection | DOAJ |
| description | The CRISPR/nCas9-based cytosine base editors (CBEs) and adenine base editors (ABEs) are capable of catalyzing C•G to T•A or A•T to G•C conversions, respectively, and have become new, powerful tools for achieving precise genetic changes in a wide range of organisms. These base editors hold great promise for correcting pathogenic mutations and for being used for therapeutic applications. However, the recognition of cognate DNA sequences near their target sites can cause severe off-target effects that greatly limit their clinical applications, and this is an urgent problem that needs to be resolved for base editing systems. The recently discovered phage-derived proteins, anti-CRISPRs, which can suppress the natural CRISPR nuclease activity, may be able to ameliorate the off-target effects of base editing systems. Here, we confirm for the first time that AcrIIA2, AcrIIA4, and AcrIIA5 efficiently inhibit base editing systems in human cells. In particular, AcrIIA5 has a significant inhibitory effect on all base editing variant systems tested in our study. We further show that the off-target effects of BE3 and ABE7.10 were significantly reduced in AcrIIA5 treated cells. This study suggests that AcrIIA5 should be widely used for the precise control of base editing and to thoroughly “shut off” nuclease activity of both CBE and ABE systems. |
| first_indexed | 2024-03-10T18:10:57Z |
| format | Article |
| id | doaj.art-3f31e74f15c641469f9b82c48664acb1 |
| institution | Directory Open Access Journal |
| issn | 2073-4409 |
| language | English |
| last_indexed | 2024-03-10T18:10:57Z |
| publishDate | 2020-07-01 |
| publisher | MDPI AG |
| record_format | Article |
| series | Cells |
| spelling | doaj.art-3f31e74f15c641469f9b82c48664acb12023-11-20T08:05:25ZengMDPI AGCells2073-44092020-07-0198178610.3390/cells9081786AcrIIA5 Suppresses Base Editors and Reduces Their Off-Target EffectsMingming Liang0Tingting Sui1Zhiquan Liu2Mao Chen3Hongmei Liu4Huanhuan Shan5Liangxue Lai6Zhanjun Li7Key Laboratory of Zoonosis Research, Ministry of Education, Jilin University, Changchun 130062, ChinaKey Laboratory of Zoonosis Research, Ministry of Education, Jilin University, Changchun 130062, ChinaKey Laboratory of Zoonosis Research, Ministry of Education, Jilin University, Changchun 130062, ChinaKey Laboratory of Zoonosis Research, Ministry of Education, Jilin University, Changchun 130062, ChinaKey Laboratory of Zoonosis Research, Ministry of Education, Jilin University, Changchun 130062, ChinaKey Laboratory of Zoonosis Research, Ministry of Education, Jilin University, Changchun 130062, ChinaKey Laboratory of Zoonosis Research, Ministry of Education, Jilin University, Changchun 130062, ChinaKey Laboratory of Zoonosis Research, Ministry of Education, Jilin University, Changchun 130062, ChinaThe CRISPR/nCas9-based cytosine base editors (CBEs) and adenine base editors (ABEs) are capable of catalyzing C•G to T•A or A•T to G•C conversions, respectively, and have become new, powerful tools for achieving precise genetic changes in a wide range of organisms. These base editors hold great promise for correcting pathogenic mutations and for being used for therapeutic applications. However, the recognition of cognate DNA sequences near their target sites can cause severe off-target effects that greatly limit their clinical applications, and this is an urgent problem that needs to be resolved for base editing systems. The recently discovered phage-derived proteins, anti-CRISPRs, which can suppress the natural CRISPR nuclease activity, may be able to ameliorate the off-target effects of base editing systems. Here, we confirm for the first time that AcrIIA2, AcrIIA4, and AcrIIA5 efficiently inhibit base editing systems in human cells. In particular, AcrIIA5 has a significant inhibitory effect on all base editing variant systems tested in our study. We further show that the off-target effects of BE3 and ABE7.10 were significantly reduced in AcrIIA5 treated cells. This study suggests that AcrIIA5 should be widely used for the precise control of base editing and to thoroughly “shut off” nuclease activity of both CBE and ABE systems.https://www.mdpi.com/2073-4409/9/8/1786base editorAnti-CRISPRoff-target effectsmammalian cells |
| spellingShingle | Mingming Liang Tingting Sui Zhiquan Liu Mao Chen Hongmei Liu Huanhuan Shan Liangxue Lai Zhanjun Li AcrIIA5 Suppresses Base Editors and Reduces Their Off-Target Effects Cells base editor Anti-CRISPR off-target effects mammalian cells |
| title | AcrIIA5 Suppresses Base Editors and Reduces Their Off-Target Effects |
| title_full | AcrIIA5 Suppresses Base Editors and Reduces Their Off-Target Effects |
| title_fullStr | AcrIIA5 Suppresses Base Editors and Reduces Their Off-Target Effects |
| title_full_unstemmed | AcrIIA5 Suppresses Base Editors and Reduces Their Off-Target Effects |
| title_short | AcrIIA5 Suppresses Base Editors and Reduces Their Off-Target Effects |
| title_sort | acriia5 suppresses base editors and reduces their off target effects |
| topic | base editor Anti-CRISPR off-target effects mammalian cells |
| url | https://www.mdpi.com/2073-4409/9/8/1786 |
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