Lentiviral Vectors Construction for Optogenetic

Lentivirus (LV) has a single stranded RNA genome with envelope and belongs to the retrovirus family. After transduction, they are stably integrated into the host chromosome to achieve permanent expression and passed on to the offspring of mitotic precursors. Therefore, LVs are suitable for mitotic and post mitotic cell types, and they have been successfully used to transduce stem cells. In post mitotic cells, LV vectors are randomly integrated, and integration occurs preferentially at active gene sites in mitotic cells. Compared with early gene delivery vectors (such as adenoviruses containing residual virus genes), LV has little or no inflammation or immune response.

Creative BioMart provides a simple and reliable method to produce high-quality LVs Vectors in a short time using ready-made materials.

Viral Vector Selection: LVs or AAV?

The most widely used vectors in optogenetics are LV and AAV.

One of the primary considerations in the selection of vector is whether the target cell type is mitotic or post mitotic.

LVS are stably integrated into the host genome, so they are suitable for both types, while AAVS are still free, so they can not support stable and long-term expression in dividing cells.

A major challenge of virus targeting is that the genome size contained in the virus capsid is limited. LV can package larger gene fragments up to 9 KB, compared with the AAV system with an upper limit of 4.7 KB.

In addition, AAV is more effective in tissue transmission than LV because they are smaller and can spread further from the injection site. Therefore, AAV is recommended when greater regional coverage is required, and LV is preferred when greater spatial specificity is required, such as when it is required for subdomains of structures (such as CA1 region of hippocampus).

Available Promoters at Creative BioMart

Promoter (serotype)	Size	Organism	Cell-Type Specificity
EF1α	1.2 kb	Rat, mouse	Neuron-specific only in LV
CMV	0.6 kb	Rat, mouse	Nonspecific (8.6% glia expressing transgene)
Human SynapsinI (hSynI)	0.5 kb	Rat, mouse	Panneuronal, but a tropism for excitatory cells in LV
CaMKIIα	1.3 kb	Macaque, rat, mouse	Excitatory neurons in cortex and hippocampus
hGFAP	2.2 kb	Rat, mouse	Astrocytes
TPH-2	2 kb	Rat	Raphe serotonergic neurons

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