The Lenti CRISPR Non-Target Control#2 Transduction Particles (LV04 vector) include a gRNA sequence that does not target known human, mouse and rat genes. The non-target control#2 transduction particles are useful as a negative control in experiments using CRISPR lentiviral clones and for cell types that are not amenable to transfection. The Lenti CRISPR Non-Target Control#2 uses a dual-component system consisting of U6-driven guide RNA (non-target#2) and hPGK-driven Puromycin resistance cassette and Blue Fluorescence Protein for fluorophorescence. This control requires Cas9 be delivered by a separate vector.
Lentiviral-based particles permit efficient infection and integration of the construct into differentiated and non-dividing cells, such as neurons and dendritic cells, overcoming low transfection and integration difficulties when using these cell lines. Self-inactivating replication incompetent viral particles are produced in packaging cells (HEK293T) by co-transfection with compatible packaging and envelope plasmids.