PUBLICATION
Mini-III RNase-based dual-color system for in vivo mRNA tracking
- Authors
- Zhang, L., Chen, L., Chen, J., Shen, W., Meng, A.
- ID
- ZDB-PUB-201024-5
- Date
- 2020
- Source
- Development (Cambridge, England) 147(22): (Journal)
- Registered Authors
- Meng, Anming
- Keywords
- DsRNA, Embryo, Mini-III RNase, RNA imaging, Zebrafish
- MeSH Terms
-
- Animals
- Bacterial Proteins/genetics
- Bacterial Proteins/metabolism*
- Fluorescein*/chemistry
- Fluorescein*/pharmacology
- Microscopy, Fluorescence
- RNA, Antisense*/chemistry
- RNA, Antisense*/pharmacology
- RNA, Messenger/genetics
- RNA, Messenger/metabolism*
- Ribonuclease III/genetics
- Ribonuclease III/metabolism*
- Staphylococcus epidermidis*/enzymology
- Staphylococcus epidermidis*/genetics
- Zebrafish/genetics
- Zebrafish/metabolism*
- PubMed
- 33093152 Full text @ Development
Citation
Zhang, L., Chen, L., Chen, J., Shen, W., Meng, A. (2020) Mini-III RNase-based dual-color system for in vivo mRNA tracking. Development (Cambridge, England). 147(22):.
Abstract
Mini-III RNase (mR3), a member of RNase III endonuclease family, can bind to and cleave double-stranded RNAs (dsRNAs). Inactive mR3 protein without the α5β-α6 loop loses the dsRNA cleavage activity, but retains dsRNA binding activity. Here, we establish an inactive mR3-based, non-engineered mR3/dsRNA system for RNA tracking in zebrafish embryos. In vitro binding experiments show that, inactive Staphylococcus epidermidis mR3 (dSmR3) protein possesses the highest binding affinity with dsRNAs among mR3s from other related species, and its binding property is retained in zebrafish embryos. Combined with a fluorescein-labeled antisense RNA probe recognizing the target mRNAs, dSmR3 tagged with an NLS and a fluorescent protein could allow visualizing the dynamics of endogenous target mRNAs. The dSmR3/antisense probe dual-color system provides a new approach to track non-engineered RNAs in real-time, which would help understand how endogenous RNAs dynamically move during embryonic development.
Genes / Markers
Expression
Phenotype
Mutations / Transgenics
Human Disease / Model
Sequence Targeting Reagents
Fish
Orthology
Engineered Foreign Genes
Mapping