PUBLICATION
Targeted RNA Knockdown by a Type III CRISPR-Cas Complex in Zebrafish
- Authors
- Fricke, T., Smalakyte, D., Lapinski, M., Pateria, A., Weige, C., Pastor, M., Kolano, A., Winata, C., Siksnys, V., Tamulaitis, G., Bochtler, M.
- ID
- ZDB-PUB-200825-10
- Date
- 2020
- Source
- The CRISPR journal 3: 299-313 (Journal)
- Registered Authors
- Bochtler, Matthias, Winata, Cecilia Lanny
- Keywords
- none
- Datasets
- GEO:GSE146852
- MeSH Terms
-
- RNA, Messenger/metabolism
- Animals, Genetically Modified
- Streptococcus thermophilus/enzymology
- CRISPR-Cas Systems*
- Gene Editing/methods*
- Animals
- Clustered Regularly Interspaced Short Palindromic Repeats
- RNA Stability*
- Zebrafish/genetics*
- PubMed
- 32833532 Full text @ CRISPR J
Citation
Fricke, T., Smalakyte, D., Lapinski, M., Pateria, A., Weige, C., Pastor, M., Kolano, A., Winata, C., Siksnys, V., Tamulaitis, G., Bochtler, M. (2020) Targeted RNA Knockdown by a Type III CRISPR-Cas Complex in Zebrafish. The CRISPR journal. 3:299-313.
Abstract
RNA interference is a powerful experimental tool for RNA knockdown, but not all organisms are amenable. Here, we provide a proof of principle demonstration that a type III Csm effector complex can be used for programmable mRNA transcript degradation in eukaryotes. In zebrafish, Streptococcus thermophilus Csm complex (StCsm) proved effective for knockdown of maternally expressed EGFP in germ cells of Tg(ddx4:ddx4-EGFP) fish. It also led to significant, albeit less drastic, fluorescence reduction at one day postfertilization in Tg(myl7:GFP) and Tg(fli1:EGFP) fish that express EGFP zygotically. StCsm targeted against the endogenous tdgf1 elicited the characteristic one-eyed phenotype with greater than 50% penetrance, and hence with similar efficiency to morpholino-mediated knockdown. We conclude that Csm-mediated knockdown is very efficient for maternal transcripts and can also be used for mixed maternal/early zygotic and early zygotic transcripts, in some cases reaching comparable efficiency to morpholino-based knockdown without significant off-target effects.
Genes / Markers
Expression
Phenotype
Mutations / Transgenics
Human Disease / Model
Sequence Targeting Reagents
Fish
Orthology
Engineered Foreign Genes
Mapping