Abstract
A microinjection system for gene delivery to a planarian was presented with materials widely used by manufacturers. The system consists of a nanoliter droplet generator/injector and a planarian holder. Glass capillary needles were used to consistently generate droplets and to inject droplets into a planarian. The holder provides a low-temperature environment that immobilizes the planarian for injection. Our system was tested and showed successful injections of microbeads and droplets with double-stranded RNA into the planarian. The results demonstrated the capability of our system as an alternative for gene delivery for studying gene functions in planarians or other living objects for regenerative medicine studies.
Highlights
Planarians, well-known flatworms that can regenerate any missing part of their bodies, are perhaps one of the most important species in regenerative medicine, which holds great promise in treating physical illnesses [1,2,3,4]
While researchers decipher the planarian regenerative mechanism, genetic tools, such as RNA interference (RNAi), in which small RNAs are employed for inhibiting translation or gene expression, are most-frequently used [6]
When the red dye was injected into mineral oil, a droplet was formed, departed from the needle tip, and sank downward onto the petri-dish in the bottom
Summary
Planarians, well-known flatworms that can regenerate any missing part of their bodies, are perhaps one of the most important species in regenerative medicine, which holds great promise in treating physical illnesses [1,2,3,4]. To perform RNAi, gene delivery is usually required. An appropriate and effective technique of gene delivery is vital in the study of planarian regeneration mechanisms. One common gene-delivery method for planarians is ingestion, whereby planarians are fed with genetic-substance-mixed food [6,7]. This method, is not applicable for studying the functions of the genes related to starvation conditions. It is difficult for planarians to eat the same amount of food mixed with genetic-substance every time, so the dosage of ingested DNA or RNA may not be consistent. A reliable gene delivery method that can be applied to planarians is required
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