Meganuclease-Based Artificial Transcription Factors
| Autor: | Hiroko Abe, Mitsuo Oshimura, Shingo Suzuki, Yoshihiro Nakajima, Takanori Miki, Ryuichiro Miura, Ken-ichi Ohta, Yasuhiro Kazuki, Hajime Shigeto, Anna Kawai |
|---|---|
| Rok vydání: | 2020 |
| Předmět: |
Transcriptional Activation
Biomedical Engineering Artificial transcription factor Gene Expression Computational biology Transfection Biochemistry Genetics and Molecular Biology (miscellaneous) Homing endonuclease Mice chemistry.chemical_compound Genes Reporter Cell Line Tumor Cricetinae Animals Humans Gene Regulatory Networks Cell Engineering Transcription factor Nuclease Reporter gene Receptors Chimeric Antigen biology DNA General Medicine DNA-binding domain Fibroblasts Endonucleases DNA-Binding Proteins HEK293 Cells chemistry Meganuclease biology.protein Transcriptome Transcription Factors |
| Zdroj: | ACS Synthetic Biology. 9:2679-2691 |
| ISSN: | 2161-5063 |
| DOI: | 10.1021/acssynbio.0c00083 |
| Popis: | Embedding middle-scale artificial gene networks in live mammalian cells is one of the most important future goals for cell engineering. However, the applications of the highly orthogonal and conventional artificial transcription factors currently available are limited. In this study, we present a scalable pipeline to produce artificial transcription factors based on homing endonucleases, also known as meganucleases. The introduction of mutations at critical sites for nuclease activity renders these homing endonucleases a simple but highly specific DNA binding domain for their specific DNA target. The introduction of inactivated meganucleases linked to transcriptional activator domains strongly induced reporter gene expression, while their fusion to transcriptional repressor domains suppressed them. In addition, we show that inactivated meganuclease-based transcription factors could be embedded in the synthetic membrane receptor synNotch and used to construct synthetic circuits. These results suggest that inactivated meganucleases are useful DNA-binding domains for the construction of synthetic transcription factors in mammalian cells. |
| Databáze: | OpenAIRE |
| Externí odkaz: |
|