ACBE, a new base editor for simultaneous C-to-T and A-to-G substitutions in mammalian systems
Abstract Background Many favorable traits of crops and livestock and human genetic diseases arise from multiple single nucleotide polymorphisms or multiple point mutations with heterogeneous base substitutions at the same locus. Current cytosine or adenine base editors can only accomplish C-to-T (G-...
| Main Authors: | , , , , , , , , , , , , , , , , , , , , |
|---|---|
| Format: | Article |
| Language: | English |
| Published: |
BMC
2020-09-01
|
| Series: | BMC Biology |
| Subjects: | |
| Online Access: | http://link.springer.com/article/10.1186/s12915-020-00866-5 |
| _version_ | 1818806583412391936 |
|---|---|
| author | Jingke Xie Xingyun Huang Xia Wang Shixue Gou Yanhui Liang Fangbing Chen Nan Li Zhen Ouyang Quanjun Zhang Weikai Ge Qin Jin Hui Shi Zhenpeng Zhuang Xiaozhu Zhao Meng Lian Jiaowei Wang Yinghua Ye Longquan Quan Han Wu Kepin Wang Liangxue Lai |
| author_facet | Jingke Xie Xingyun Huang Xia Wang Shixue Gou Yanhui Liang Fangbing Chen Nan Li Zhen Ouyang Quanjun Zhang Weikai Ge Qin Jin Hui Shi Zhenpeng Zhuang Xiaozhu Zhao Meng Lian Jiaowei Wang Yinghua Ye Longquan Quan Han Wu Kepin Wang Liangxue Lai |
| author_sort | Jingke Xie |
| collection | DOAJ |
| description | Abstract Background Many favorable traits of crops and livestock and human genetic diseases arise from multiple single nucleotide polymorphisms or multiple point mutations with heterogeneous base substitutions at the same locus. Current cytosine or adenine base editors can only accomplish C-to-T (G-to-A) or A-to-G (T-to-C) substitutions in the windows of target genomic sites of organisms; therefore, there is a need to develop base editors that can simultaneously achieve C-to-T and A-to-G substitutions at the targeting site. Results In this study, a novel fusion adenine and cytosine base editor (ACBE) was generated by fusing a heterodimer of TadA (ecTadAWT/*) and an activation-induced cytidine deaminase (AID) to the N- and C-terminals of Cas9 nickase (nCas9), respectively. ACBE could simultaneously induce C-to-T and A-to-G base editing at the same target site, which were verified in HEK293-EGFP reporter cell line and 45 endogenous gene loci of HEK293 cells. Moreover, the ACBE could accomplish simultaneous point mutations of C-to-T and A-to-G in primary somatic cells (mouse embryonic fibroblasts and porcine fetal fibroblasts) in an applicable efficiency. Furthermore, the spacer length of sgRNA and the length of linker could influence the dual base editing activity, which provided a direction to optimize the ACBE system. Conclusion The newly developed ACBE would expand base editor toolkits and should promote the generation of animals and the gene therapy of genetic diseases with heterogeneous point mutations. |
| first_indexed | 2024-12-18T19:12:05Z |
| format | Article |
| id | doaj.art-5974bfd9c0c64e52a24cfd1869de793b |
| institution | Directory Open Access Journal |
| issn | 1741-7007 |
| language | English |
| last_indexed | 2024-12-18T19:12:05Z |
| publishDate | 2020-09-01 |
| publisher | BMC |
| record_format | Article |
| series | BMC Biology |
| spelling | doaj.art-5974bfd9c0c64e52a24cfd1869de793b2022-12-21T20:56:14ZengBMCBMC Biology1741-70072020-09-0118111410.1186/s12915-020-00866-5ACBE, a new base editor for simultaneous C-to-T and A-to-G substitutions in mammalian systemsJingke Xie0Xingyun Huang1Xia Wang2Shixue Gou3Yanhui Liang4Fangbing Chen5Nan Li6Zhen Ouyang7Quanjun Zhang8Weikai Ge9Qin Jin10Hui Shi11Zhenpeng Zhuang12Xiaozhu Zhao13Meng Lian14Jiaowei Wang15Yinghua Ye16Longquan Quan17Han Wu18Kepin Wang19Liangxue Lai20CAS Key Laboratory of Regenerative Biology, Guangdong Provincial Key Laboratory of Stem Cell and Regenerative Medicine, Guangzhou Institutes of Biomedicine and Health, Chinese Academy of SciencesCAS Key Laboratory of Regenerative Biology, Guangdong Provincial Key Laboratory of Stem Cell and Regenerative Medicine, Guangzhou Institutes of Biomedicine and Health, Chinese Academy of SciencesCAS Key Laboratory of Regenerative Biology, Guangdong Provincial Key Laboratory of Stem Cell and Regenerative Medicine, Guangzhou Institutes of Biomedicine and Health, Chinese Academy of SciencesCAS Key Laboratory of Regenerative Biology, Guangdong Provincial Key Laboratory of Stem Cell and Regenerative Medicine, Guangzhou Institutes of Biomedicine and Health, Chinese Academy of SciencesCAS Key Laboratory of Regenerative Biology, Guangdong Provincial Key Laboratory of Stem Cell and Regenerative Medicine, Guangzhou Institutes of Biomedicine and Health, Chinese Academy of SciencesCAS Key Laboratory of Regenerative Biology, Guangdong Provincial Key Laboratory of Stem Cell and Regenerative Medicine, Guangzhou Institutes of Biomedicine and Health, Chinese Academy of SciencesCAS Key Laboratory of Regenerative Biology, Guangdong Provincial Key Laboratory of Stem Cell and Regenerative Medicine, Guangzhou Institutes of Biomedicine and Health, Chinese Academy of SciencesCAS Key Laboratory of Regenerative Biology, Guangdong Provincial Key Laboratory of Stem Cell and Regenerative Medicine, Guangzhou Institutes of Biomedicine and Health, Chinese Academy of SciencesCAS Key Laboratory of Regenerative Biology, Guangdong Provincial Key Laboratory of Stem Cell and Regenerative Medicine, Guangzhou Institutes of Biomedicine and Health, Chinese Academy of SciencesCAS Key Laboratory of Regenerative Biology, Guangdong Provincial Key Laboratory of Stem Cell and Regenerative Medicine, Guangzhou Institutes of Biomedicine and Health, Chinese Academy of SciencesCAS Key Laboratory of Regenerative Biology, Guangdong Provincial Key Laboratory of Stem Cell and Regenerative Medicine, Guangzhou Institutes of Biomedicine and Health, Chinese Academy of SciencesCAS Key Laboratory of Regenerative Biology, Guangdong Provincial Key Laboratory of Stem Cell and Regenerative Medicine, Guangzhou Institutes of Biomedicine and Health, Chinese Academy of SciencesCAS Key Laboratory of Regenerative Biology, Guangdong Provincial Key Laboratory of Stem Cell and Regenerative Medicine, Guangzhou Institutes of Biomedicine and Health, Chinese Academy of SciencesCAS Key Laboratory of Regenerative Biology, Guangdong Provincial Key Laboratory of Stem Cell and Regenerative Medicine, Guangzhou Institutes of Biomedicine and Health, Chinese Academy of SciencesCAS Key Laboratory of Regenerative Biology, Guangdong Provincial Key Laboratory of Stem Cell and Regenerative Medicine, Guangzhou Institutes of Biomedicine and Health, Chinese Academy of SciencesCAS Key Laboratory of Regenerative Biology, Guangdong Provincial Key Laboratory of Stem Cell and Regenerative Medicine, Guangzhou Institutes of Biomedicine and Health, Chinese Academy of SciencesCAS Key Laboratory of Regenerative Biology, Guangdong Provincial Key Laboratory of Stem Cell and Regenerative Medicine, Guangzhou Institutes of Biomedicine and Health, Chinese Academy of SciencesCAS Key Laboratory of Regenerative Biology, Guangdong Provincial Key Laboratory of Stem Cell and Regenerative Medicine, Guangzhou Institutes of Biomedicine and Health, Chinese Academy of SciencesCAS Key Laboratory of Regenerative Biology, Guangdong Provincial Key Laboratory of Stem Cell and Regenerative Medicine, Guangzhou Institutes of Biomedicine and Health, Chinese Academy of SciencesCAS Key Laboratory of Regenerative Biology, Guangdong Provincial Key Laboratory of Stem Cell and Regenerative Medicine, Guangzhou Institutes of Biomedicine and Health, Chinese Academy of SciencesCAS Key Laboratory of Regenerative Biology, Guangdong Provincial Key Laboratory of Stem Cell and Regenerative Medicine, Guangzhou Institutes of Biomedicine and Health, Chinese Academy of SciencesAbstract Background Many favorable traits of crops and livestock and human genetic diseases arise from multiple single nucleotide polymorphisms or multiple point mutations with heterogeneous base substitutions at the same locus. Current cytosine or adenine base editors can only accomplish C-to-T (G-to-A) or A-to-G (T-to-C) substitutions in the windows of target genomic sites of organisms; therefore, there is a need to develop base editors that can simultaneously achieve C-to-T and A-to-G substitutions at the targeting site. Results In this study, a novel fusion adenine and cytosine base editor (ACBE) was generated by fusing a heterodimer of TadA (ecTadAWT/*) and an activation-induced cytidine deaminase (AID) to the N- and C-terminals of Cas9 nickase (nCas9), respectively. ACBE could simultaneously induce C-to-T and A-to-G base editing at the same target site, which were verified in HEK293-EGFP reporter cell line and 45 endogenous gene loci of HEK293 cells. Moreover, the ACBE could accomplish simultaneous point mutations of C-to-T and A-to-G in primary somatic cells (mouse embryonic fibroblasts and porcine fetal fibroblasts) in an applicable efficiency. Furthermore, the spacer length of sgRNA and the length of linker could influence the dual base editing activity, which provided a direction to optimize the ACBE system. Conclusion The newly developed ACBE would expand base editor toolkits and should promote the generation of animals and the gene therapy of genetic diseases with heterogeneous point mutations.http://link.springer.com/article/10.1186/s12915-020-00866-5Adenine and cytosine base editor (ACBE)Simultaneous C-to-T and A-to-G conversionsMammalian systems |
| spellingShingle | Jingke Xie Xingyun Huang Xia Wang Shixue Gou Yanhui Liang Fangbing Chen Nan Li Zhen Ouyang Quanjun Zhang Weikai Ge Qin Jin Hui Shi Zhenpeng Zhuang Xiaozhu Zhao Meng Lian Jiaowei Wang Yinghua Ye Longquan Quan Han Wu Kepin Wang Liangxue Lai ACBE, a new base editor for simultaneous C-to-T and A-to-G substitutions in mammalian systems BMC Biology Adenine and cytosine base editor (ACBE) Simultaneous C-to-T and A-to-G conversions Mammalian systems |
| title | ACBE, a new base editor for simultaneous C-to-T and A-to-G substitutions in mammalian systems |
| title_full | ACBE, a new base editor for simultaneous C-to-T and A-to-G substitutions in mammalian systems |
| title_fullStr | ACBE, a new base editor for simultaneous C-to-T and A-to-G substitutions in mammalian systems |
| title_full_unstemmed | ACBE, a new base editor for simultaneous C-to-T and A-to-G substitutions in mammalian systems |
| title_short | ACBE, a new base editor for simultaneous C-to-T and A-to-G substitutions in mammalian systems |
| title_sort | acbe a new base editor for simultaneous c to t and a to g substitutions in mammalian systems |
| topic | Adenine and cytosine base editor (ACBE) Simultaneous C-to-T and A-to-G conversions Mammalian systems |
| url | http://link.springer.com/article/10.1186/s12915-020-00866-5 |
| work_keys_str_mv | AT jingkexie acbeanewbaseeditorforsimultaneousctotandatogsubstitutionsinmammaliansystems AT xingyunhuang acbeanewbaseeditorforsimultaneousctotandatogsubstitutionsinmammaliansystems AT xiawang acbeanewbaseeditorforsimultaneousctotandatogsubstitutionsinmammaliansystems AT shixuegou acbeanewbaseeditorforsimultaneousctotandatogsubstitutionsinmammaliansystems AT yanhuiliang acbeanewbaseeditorforsimultaneousctotandatogsubstitutionsinmammaliansystems AT fangbingchen acbeanewbaseeditorforsimultaneousctotandatogsubstitutionsinmammaliansystems AT nanli acbeanewbaseeditorforsimultaneousctotandatogsubstitutionsinmammaliansystems AT zhenouyang acbeanewbaseeditorforsimultaneousctotandatogsubstitutionsinmammaliansystems AT quanjunzhang acbeanewbaseeditorforsimultaneousctotandatogsubstitutionsinmammaliansystems AT weikaige acbeanewbaseeditorforsimultaneousctotandatogsubstitutionsinmammaliansystems AT qinjin acbeanewbaseeditorforsimultaneousctotandatogsubstitutionsinmammaliansystems AT huishi acbeanewbaseeditorforsimultaneousctotandatogsubstitutionsinmammaliansystems AT zhenpengzhuang acbeanewbaseeditorforsimultaneousctotandatogsubstitutionsinmammaliansystems AT xiaozhuzhao acbeanewbaseeditorforsimultaneousctotandatogsubstitutionsinmammaliansystems AT menglian acbeanewbaseeditorforsimultaneousctotandatogsubstitutionsinmammaliansystems AT jiaoweiwang acbeanewbaseeditorforsimultaneousctotandatogsubstitutionsinmammaliansystems AT yinghuaye acbeanewbaseeditorforsimultaneousctotandatogsubstitutionsinmammaliansystems AT longquanquan acbeanewbaseeditorforsimultaneousctotandatogsubstitutionsinmammaliansystems AT hanwu acbeanewbaseeditorforsimultaneousctotandatogsubstitutionsinmammaliansystems AT kepinwang acbeanewbaseeditorforsimultaneousctotandatogsubstitutionsinmammaliansystems AT liangxuelai acbeanewbaseeditorforsimultaneousctotandatogsubstitutionsinmammaliansystems |