LentiviraL Vector Design for Mitochondria Localization
Overview of Mitochondria Localization
Mitochondrial localization is a technique for studying mitochondrial functioning by targeting specific proteins or compounds in different subregions of the mitochondria. This method involves the fusing of mitochondrial localization signal (MLS) or mitochondrial targeting sequence (MTS) with the target protein or molecule, which allows for precise localization to various compartments inside the mitochondria, such as the matrix, inner membrane, and outer membrane. MTS is a short peptide, 15-70 amino acids long, with positively charged basic residues that drive protein transport to the mitochondria.
Lentiviral Vector for Mitochondria Localization
MTS-based mitochondrial targeting techniques were applied to fusion into viral vectors for targeted delivery. Creative Biolabs created an MTS-conjugated lentiviral vector (LV) that inserts the MTS signal into the transfer plasmid and transfects it into 293T cells using the calcium phosphate technique for lentiviral packaging. We offer several common LVs for mitochondria localization.
- LV-mito-GFP uses green fluorescent protein (GFP) to visualize mitochondria, facilitating dynamic studies of shape and motility.
- LV-mtDsRed expresses the red fluorescent protein DsRed coupled with a mitochondrial targeting sequence, allowing for high-resolution imaging of mitochondria in live cells.
- LV-mito-mCherry uses the mCherry fluorescent protein for multi-channel imaging and colocalization research.
Fig.1 Lentiviral vector plasmid pLV-Mito-GFP.1
Applications of LV for Mitochondria Localization
LV with mitochondrial colocalization is appropriate for establishing stably transduced cell lines. The primary mitochondrial localization stably transduced cell lines we can offer are:
- HeLa Cell Line
Stably transfected with the LV-mtDsRed, the HeLa cell line is a fluorescent tool for mitochondrial localization. This fusion protein, which is intended to be localized in mitochondria, produces red fluorescence when illuminated. HeLa cells are widely available and easy to cultivate, making them popular for research on mitochondrial dynamics and functions.
- HEK293 Cell Line
Similar to HeLa cells, the HEK293 cell line allows for easy cultivation and transfection. These cells generate green fluorescence when illuminated by mitochondria thanks to stable transfection LV-mito-GFP. This fluorescent allows for high-precision observations of mitochondrial morphology, mobility, and interactions in HEK293 cells.
- Neuro-2a Cell Line
The Neuro-2a cell line, also a neuroblastoma cell line, can emit red fluorescence under mitochondrial illumination after transfection of LV-mito-mCherry. It is to be used to study the relationship between mitochondria and neurodegenerative diseases.
In the above cell lines, stable transfection of LVs can make the mitochondrial fluorescent PCR label stably concentrated inside the mitochondria, making it easier to observe and analyze mitochondria. Different cell lines and labeling methods are suitable for different research questions, so researchers should choose the most suitable mitochondrial localization stably transduced cell line according to their own needs.
Creative Biolabs is dedicated to offering high-quality LVs for mitochondria localization services to customers worldwide. For more extensive information, please contact us and request a price. We will contact you within 24 hours.
Reference
- Kitay, Brandon M., et al. "Mislocalization of neuronal mitochondria reveals regulation of Wallerian degeneration and NMNAT/WLDS-mediated axon protection independent of axonal mitochondria." Human molecular genetics 22.8 (2013): 1601-1614.
