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In transgenic research, selection markers have greatly facilitated the generation of transgenic animals. A prerequisite for a suitable selection marker to be used along with a test gene of interest is that the marker should not affect the phenotype of interest in transformed animals. One of the most common selection markers used in  C .  elegans  transgenic approaches is the  rol-6  co-injection marker, which induces a behavioral roller phenotype due to a cuticle defect but is not known to have other side effects. However, we found that the  rol-6  co-injection marker can cause expression of GFP in the test sequence in a male-specific interneuron called CP09. We found that the  rol-6  gene sequence included in the marker plasmid is responsible for this unwanted expression. Accordingly, the use of the  rol-6  co-injection marker is not recommended when researchers intend to examine precise expression or perform functional studies especially targeting male  C .  elegans  neurons. The  rol-6  sequence region we identified can be used to drive a specific expression in CP09 neuron for future research.

Expressional artifact caused by a co-injection marker rol-6 in C. elegans