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

Autoři: HoYong Jin aff001;  Scott W. Emmons aff002;  Byunghyuk Kim aff001
Působiště autorů: Department of Life Science, Dongguk University-Seoul, Goyang, Republic of Korea aff001;  Department of Genetics and Dominick P. Purpura Department of Neuroscience, Albert Einstein College of Medicine, Bronx, New York, United States of America aff002
Vyšlo v časopise: PLoS ONE 14(12)
Kategorie: Research Article
prolekare.web.journal.doi_sk: 10.1371/journal.pone.0224533


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.

Klíčová slova:

3' UTR – Caenorhabditis elegans – Gene expression – Genetically modified animals – Homologous recombination – Neurons – Plasmid construction – Plasmid vectors


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2019 Číslo 12