Dear Mounir El Hankouri ,

If you’re looking to label TPM3.1 in live cells, here are a couple of options to consider:

1. Sir-Dye Labeling: Currently, there isn't a Sir-Dye specific to TPM3.1, as Sir-Dyes are generally used for broader targets like cytoskeletal components (e.g., SiR-Actin for actin filaments, SiR-Tubulin for microtubules). Since TPM3.1 is a tropomyosin isoform, Sir-Actin may indirectly help visualize its binding along actin filaments, but it won’t specifically target TPM3.1 itself.

2. Plasmid for IVT (mRNA) Tagging: For direct labeling, you may want to consider using a TPM3.1 construct tagged with GFP or RFP. There are commercially available plasmids for tropomyosin isoforms tagged with fluorescent proteins; some vendors offer custom tagged mRNA constructs as well, suitable for in vitro transcription. For example, you could use a TPM3.1-GFP or RFP tagged plasmid for direct expression and fluorescent visualization. If you don’t find a TPM3.1-specific construct, creating a custom TPM3.1-GFP/RFP fusion via cloning could also be a viable route.

Dear Mounir,

Currently, there is no commercially available Sir-dye specific to TPM3.1. However, a plasmid suitable for in vitro transcription of TPM3 is accessible, allowing you to prepare RNA for injection experiments. For instance, the DNASU Plasmid Repository offers a human TPM3 gene plasmid in an in vitro expression vector (pANT7_cGST), which may be suitable for generating transcripts for experimental applications (https://atm.amegroups.org/article/view/77642/html, chrome-extension://efaidnbmnnnibpcajpcglclefindmkaj/https://www.insidehopkinsmedicine.org/mbg/Publications/2013/Green/Green_InVitro_Transcription_Plasmid_PCR_Amplified_DNA.pdf)

For in vitro transcription, you can follow protocols using T7 RNA polymerase and linearized DNA templates to generate RNA transcripts from these plasmids

Thank you so much for your suggestions ! Have a great day