Efficient multitool/multiplex gene engineering with TALE-BE
TALE base editors are a recent addition to the genome editing toolbox. These molecular tools are fusions of a transcription activator-like effector domain (TALE), split-DddA deaminase halves, and an uracil glycosylase inhibitor (UGI) that have the distinct ability to directly edit double strand DNA,...
| Main Authors: | , , , , , , , , |
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| Format: | Article |
| Language: | English |
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Frontiers Media S.A.
2022-11-01
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| Series: | Frontiers in Bioengineering and Biotechnology |
| Subjects: | |
| Online Access: | https://www.frontiersin.org/articles/10.3389/fbioe.2022.1033669/full |
| _version_ | 1828099055847211008 |
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| author | Alex Boyne Ming Yang Sylvain Pulicani Maria Feola Diane Tkach Robert Hong Aymeric Duclert Philippe Duchateau Alexandre Juillerat |
| author_facet | Alex Boyne Ming Yang Sylvain Pulicani Maria Feola Diane Tkach Robert Hong Aymeric Duclert Philippe Duchateau Alexandre Juillerat |
| author_sort | Alex Boyne |
| collection | DOAJ |
| description | TALE base editors are a recent addition to the genome editing toolbox. These molecular tools are fusions of a transcription activator-like effector domain (TALE), split-DddA deaminase halves, and an uracil glycosylase inhibitor (UGI) that have the distinct ability to directly edit double strand DNA, converting a cytosine (C) to a thymine (T). To dissect the editing rules of TALE-BE, we combined the screening of dozens of TALE-BE targeting nuclear genomic loci with a medium/high throughput strategy based on precise knock-in of TALE-BE target site collections into the cell genome. This latter approach allowed us to gain in depth insight of the editing rules in cellulo, while excluding confounding factors such as epigenetic and microenvironmental differences among different genomic loci. Using the knowledge gained, we designed TALE-BE targeting CD52 and achieved very high frequency of gene knock-out (up to 80% of phenotypic CD52 knock out). We further demonstrated that TALE-BE generate only insignificant levels of Indels and byproducts. Finally, we combined two molecular tools, a TALE-BE and a TALEN, for multiplex genome engineering, generating high levels of double gene knock-out (∼75%) without creation of translocations between the two targeted sites. |
| first_indexed | 2024-04-11T08:11:28Z |
| format | Article |
| id | doaj.art-5859141248d2427197bdff48d14a5260 |
| institution | Directory Open Access Journal |
| issn | 2296-4185 |
| language | English |
| last_indexed | 2024-04-11T08:11:28Z |
| publishDate | 2022-11-01 |
| publisher | Frontiers Media S.A. |
| record_format | Article |
| series | Frontiers in Bioengineering and Biotechnology |
| spelling | doaj.art-5859141248d2427197bdff48d14a52602022-12-22T04:35:21ZengFrontiers Media S.A.Frontiers in Bioengineering and Biotechnology2296-41852022-11-011010.3389/fbioe.2022.10336691033669Efficient multitool/multiplex gene engineering with TALE-BEAlex Boyne0Ming Yang1Sylvain Pulicani2Maria Feola3Diane Tkach4Robert Hong5Aymeric Duclert6Philippe Duchateau7Alexandre Juillerat8Cellectis Inc., New York, NY, United StatesCellectis Inc., New York, NY, United StatesCellectis, Paris, FranceCellectis Inc., New York, NY, United StatesCellectis Inc., New York, NY, United StatesCellectis Inc., New York, NY, United StatesCellectis, Paris, FranceCellectis, Paris, FranceCellectis Inc., New York, NY, United StatesTALE base editors are a recent addition to the genome editing toolbox. These molecular tools are fusions of a transcription activator-like effector domain (TALE), split-DddA deaminase halves, and an uracil glycosylase inhibitor (UGI) that have the distinct ability to directly edit double strand DNA, converting a cytosine (C) to a thymine (T). To dissect the editing rules of TALE-BE, we combined the screening of dozens of TALE-BE targeting nuclear genomic loci with a medium/high throughput strategy based on precise knock-in of TALE-BE target site collections into the cell genome. This latter approach allowed us to gain in depth insight of the editing rules in cellulo, while excluding confounding factors such as epigenetic and microenvironmental differences among different genomic loci. Using the knowledge gained, we designed TALE-BE targeting CD52 and achieved very high frequency of gene knock-out (up to 80% of phenotypic CD52 knock out). We further demonstrated that TALE-BE generate only insignificant levels of Indels and byproducts. Finally, we combined two molecular tools, a TALE-BE and a TALEN, for multiplex genome engineering, generating high levels of double gene knock-out (∼75%) without creation of translocations between the two targeted sites.https://www.frontiersin.org/articles/10.3389/fbioe.2022.1033669/fullgene editingbase editorsTALEt-cellscell engineering |
| spellingShingle | Alex Boyne Ming Yang Sylvain Pulicani Maria Feola Diane Tkach Robert Hong Aymeric Duclert Philippe Duchateau Alexandre Juillerat Efficient multitool/multiplex gene engineering with TALE-BE Frontiers in Bioengineering and Biotechnology gene editing base editors TALE t-cells cell engineering |
| title | Efficient multitool/multiplex gene engineering with TALE-BE |
| title_full | Efficient multitool/multiplex gene engineering with TALE-BE |
| title_fullStr | Efficient multitool/multiplex gene engineering with TALE-BE |
| title_full_unstemmed | Efficient multitool/multiplex gene engineering with TALE-BE |
| title_short | Efficient multitool/multiplex gene engineering with TALE-BE |
| title_sort | efficient multitool multiplex gene engineering with tale be |
| topic | gene editing base editors TALE t-cells cell engineering |
| url | https://www.frontiersin.org/articles/10.3389/fbioe.2022.1033669/full |
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