Tyrosinase-GCaMP6-Cherry was created by cloning mCherry after GCaMP6. MELOPS was engineered by replacing the triple HA tag in pCR3-OCA2-HA with Nectarine [Addgene 21717 (25)]. Site-directed mutagenesis was performed with Quickchange II kit (Agilent Technologies) to introduce the OCA2-V443I mutation in MELOPS.
Melanin pigments are made in skin and hair cells called melanocytes and provide color to skin and hair and protection against UV rays. Inadequate protection poses the risk of genetic mutations which can lead to skin cancer.
This plasmid allows for expression of the human tyrosinase enzyme fused to infra-red fluorescent protein (iRFP). The tyrosinase enzyme catalyzes the rate limiting step in the synthesis of melanin pigment. The iRFP allows for visualization of the protein by fluorescence microscopy. The plasmid has a CMV promoter, which is typically associated with strong expression levels in mammalian cells.
Santiago Di Pietro
Andrea Ambrosio
Steve Foster
Steve.Foster@colostate.edu
970-491-7100