Cutting edge Seminar
Speaker: Miki Imanishi (Serior Lecturer, Biofunctional Design Chemistry, Division of Biochemistry, Kyoto University Institute for Chemical Research）
Title: Design of nucleic acid binding proteins to control cellular events
Date&Time: 11 Jul. (Wed.) 2018, 12:00- 13:00
Venue: Conference Room(1F), IMEG
Abstract:
Artificial DNA or RNA binding proteins binding to desired genomic or mRNA regions are useful to edit genome sequences or control gene expressions. In addition to the recently developed CRISPR-Cas9 gene targeting system, C2H2-type zinc finger domains and TALEs (Transcription Activator-Like Effectors) have provided attractive scaffolds to design such sequence-specific DNA binding proteins. In this presentation, I would like to introduce our approaches about (1) design of artificial zinc finger proteins for manipulation of the cellular circadian clock, (2) directed evolution of TALEs with new DNA binding properties and (3) development of artificial RNA binding proteins.
In the first topic, we aimed to manipulate the mammalian circadian clock by designing zinc finger-based artificial transcription factors. We designed a transcription factor specifically bound to the regulatory region of the Period1 gene and showed that the artificial protein led to circadian phase resetting [1a]. We also constructed a simple system to manipulate gene expression patterns to be circadian [1b]. These approaches potentially open the way to understand the molecular mechanisms of circadian rhythm and to create an oscillating gene expression system. In the second topic, we performed directed evolution of a TALE repeat to specifically recognize 5mC to detect and regulate specific 5mC in living cells [2]. In the third topic, I will introduce development of artificial RNA binding proteins that can bind to specific 16-nt RNA and control a single specific mRNA [3]. Development of DNA or RNA binding proteins in a sequence specific manner would help understanding of various cellular events.
References
1a. Imanishi, M., Nakamura, A., Doi, M., Futaki, S., Okamura, H. “Resetting the Circadian Clock by an Artificial Zinc Finger Transcription Regulator” Angew. Chem. Int. Ed., 50, 9396-9399 (2011).
1b. Imanishi, M., Yamamoto, K., Yamada, H., Hirose, Y. Okamura, H., and Futaki, S. “Construction of a Rhythm Transfer System that Mimics the Cellular Clock” ACS Chem. Biol., 7, 1817-1821 (2012).
- Tsuji, S., Futaki, S., and Imanishi, M. “Sequence-Specific Recognition of Methylated DNA by an Engineered Transcription Activator-Like Effector Protein” Chem. Commun., 52, 14238-14241(2016).
- Shinoda, K., Tsuji, S., Futaki, S., and Imanishi, M. “Nested PUF Proteins: Extending Target RNA Elements for Gene Regulation” ChemBioChem, 19, 171-176 (2018).
