Pioneer Factor Improves CRISPR‐Based C‐To‐G and C‐To‐T Base Editing
Abstract Base editing events in eukaryote require a compatible chromatin environment, but there is little research on how chromatin factors contribute to the editing efficiency or window. By engineering BEs (base editors) fused with various pioneer factors, the authors found that SOX2 substantially...
| Main Authors: | , , , , , |
|---|---|
| Format: | Article |
| Language: | English |
| Published: |
Wiley
2022-09-01
|
| Series: | Advanced Science |
| Subjects: | |
| Online Access: | https://doi.org/10.1002/advs.202202957 |
| _version_ | 1797817532319531008 |
|---|---|
| author | Chao Yang Xingxiao Dong Zhenzhen Ma Bo Li Changhao Bi Xueli Zhang |
| author_facet | Chao Yang Xingxiao Dong Zhenzhen Ma Bo Li Changhao Bi Xueli Zhang |
| author_sort | Chao Yang |
| collection | DOAJ |
| description | Abstract Base editing events in eukaryote require a compatible chromatin environment, but there is little research on how chromatin factors contribute to the editing efficiency or window. By engineering BEs (base editors) fused with various pioneer factors, the authors found that SOX2 substantially increased the editing efficiency for GBE and CBE. While SoxN‐GBE (SOX2‐NH3‐GBE) improved the editing efficiency at overall cytosines of the protospacer, SoxM‐GBE/CBE (SOX2‐Middle‐GBE/CBE) enabled the higher base editing at PAM‐proximal cytosines. By separating functional domains of SOX2, the SadN‐GBE (SOX2 activation domain‐NH3‐GBE) is constructed for higher editing efficiency and SadM‐CBE for broader editing window to date. With the DNase I assay, it is also proved the increased editing efficiency is most likely associated with the induction of chromatin accessibility by SAD. Finally, SadM‐CBE is employed to introduce a stop codon in the proto‐oncogene MYC, at a locus rarely edited by previous editors with high efficiency. In this work, a new class of pioneer‐BEs is constructed by fusion of pioneer factor or its functional domains, which exhibits higher editing efficiency or broader editing window in eukaryote. |
| first_indexed | 2024-03-13T08:54:48Z |
| format | Article |
| id | doaj.art-f1d8b201363a460ba9e3c9275e834ef7 |
| institution | Directory Open Access Journal |
| issn | 2198-3844 |
| language | English |
| last_indexed | 2024-03-13T08:54:48Z |
| publishDate | 2022-09-01 |
| publisher | Wiley |
| record_format | Article |
| series | Advanced Science |
| spelling | doaj.art-f1d8b201363a460ba9e3c9275e834ef72023-05-29T04:01:39ZengWileyAdvanced Science2198-38442022-09-01926n/an/a10.1002/advs.202202957Pioneer Factor Improves CRISPR‐Based C‐To‐G and C‐To‐T Base EditingChao Yang0Xingxiao Dong1Zhenzhen Ma2Bo Li3Changhao Bi4Xueli Zhang5Tianjin Institute of Industrial Biotechnology Chinese Academy of Sciences Tianjin 300308 ChinaSchool of Biological Engineering Dalian Polytechnic University Dalian 116034 ChinaCollege of Life Sciences Nankai University Tianjin 300071 ChinaTianjin Institute of Industrial Biotechnology Chinese Academy of Sciences Tianjin 300308 ChinaTianjin Institute of Industrial Biotechnology Chinese Academy of Sciences Tianjin 300308 ChinaTianjin Institute of Industrial Biotechnology Chinese Academy of Sciences Tianjin 300308 ChinaAbstract Base editing events in eukaryote require a compatible chromatin environment, but there is little research on how chromatin factors contribute to the editing efficiency or window. By engineering BEs (base editors) fused with various pioneer factors, the authors found that SOX2 substantially increased the editing efficiency for GBE and CBE. While SoxN‐GBE (SOX2‐NH3‐GBE) improved the editing efficiency at overall cytosines of the protospacer, SoxM‐GBE/CBE (SOX2‐Middle‐GBE/CBE) enabled the higher base editing at PAM‐proximal cytosines. By separating functional domains of SOX2, the SadN‐GBE (SOX2 activation domain‐NH3‐GBE) is constructed for higher editing efficiency and SadM‐CBE for broader editing window to date. With the DNase I assay, it is also proved the increased editing efficiency is most likely associated with the induction of chromatin accessibility by SAD. Finally, SadM‐CBE is employed to introduce a stop codon in the proto‐oncogene MYC, at a locus rarely edited by previous editors with high efficiency. In this work, a new class of pioneer‐BEs is constructed by fusion of pioneer factor or its functional domains, which exhibits higher editing efficiency or broader editing window in eukaryote.https://doi.org/10.1002/advs.202202957CRISPR/Cas9base editingchromatin accessibilitypioneer factor |
| spellingShingle | Chao Yang Xingxiao Dong Zhenzhen Ma Bo Li Changhao Bi Xueli Zhang Pioneer Factor Improves CRISPR‐Based C‐To‐G and C‐To‐T Base Editing Advanced Science CRISPR/Cas9 base editing chromatin accessibility pioneer factor |
| title | Pioneer Factor Improves CRISPR‐Based C‐To‐G and C‐To‐T Base Editing |
| title_full | Pioneer Factor Improves CRISPR‐Based C‐To‐G and C‐To‐T Base Editing |
| title_fullStr | Pioneer Factor Improves CRISPR‐Based C‐To‐G and C‐To‐T Base Editing |
| title_full_unstemmed | Pioneer Factor Improves CRISPR‐Based C‐To‐G and C‐To‐T Base Editing |
| title_short | Pioneer Factor Improves CRISPR‐Based C‐To‐G and C‐To‐T Base Editing |
| title_sort | pioneer factor improves crispr based c to g and c to t base editing |
| topic | CRISPR/Cas9 base editing chromatin accessibility pioneer factor |
| url | https://doi.org/10.1002/advs.202202957 |
| work_keys_str_mv | AT chaoyang pioneerfactorimprovescrisprbasedctogandctotbaseediting AT xingxiaodong pioneerfactorimprovescrisprbasedctogandctotbaseediting AT zhenzhenma pioneerfactorimprovescrisprbasedctogandctotbaseediting AT boli pioneerfactorimprovescrisprbasedctogandctotbaseediting AT changhaobi pioneerfactorimprovescrisprbasedctogandctotbaseediting AT xuelizhang pioneerfactorimprovescrisprbasedctogandctotbaseediting |