Targeted RNA Knockdown by a Type III CRISPR-Cas Complex in Zebrafish.
|Targeted RNA Knockdown by a Type III CRISPR-Cas Complex in Zebrafish.
|Year of Publication
|Fricke, T, Smalakyte, D, Lapinski, M, Pateria, A, Weige, C, Pastor, M, Kolano, A, Winata, C, Siksnys, V, Tamulaitis, G, Bochtler, M
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, Csm complex (StCsm) proved effective for knockdown of maternally expressed in germ cells of fish. It also led to significant, albeit less drastic, fluorescence reduction at one day postfertilization in and ( fish that express zygotically. StCsm targeted against the endogenous 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.
|PubMed Central ID