Lumiere CRISPR/Cas9 Nanoparticles (Cas9/sgRNA NPs) consist of Cas9 proteins, sgRNAs (designed by customers), and granule scaffold proteins. All co-expressed elements mediate the release of Lumiere NPs from the cell membrane of producing cells through self-assembly.

Secreted Lumiere NPs can be harvested/concentrated, and then used to transduction. Once the NPs are recognized and fused into target cells, the active Cas9/sgRNA RNP cargoes would be further transported to the nucleus for effective gene editing.

How does CRISPR/Cas9/sgRNA NPs work?

Fig1. The process for packaging targeted CRISPR Cas9 RNP nanoparticles.

1. Dilute the purified target plasmid to a total volume of 750 μl and add it to the Cas9/sgRNA NPs Packaging Mix 1. Vortex the solution to ensure complete mixing.

2. Carefully add the entire contents of the mixed solution from step 1 into the Cas9/sgRNA NPs Packaging Mix 2. Gently mix the solutions without creating air bubbles.

3. Transfer the combined mixture onto a pre-labeled petri dish containing the cells of interest (such as 293T cells). Incubate the cells with the mixture under appropriate conditions as specified by your protocol.

4. After an incubation period of 48-72 hours, perform a brief centrifugation at the recommended speed and duration to pellet the cells. Carefully aspirate and collect the supernatant for further processing or analysis.

Features

. A remarkably high transfection efficiency ranging from 50% to nearly 100% has been demonstrated, even in cell types that are typically refractory to transfection.

. NPs are not constrained by the size limitations associated with Adeno-Associated Virus (AAV) vectors

. Unlike Lentiviral (LV) vectors, NPs do not integrate randomly into the genome

· Low cytotoxicity and non-effect on cell viability and growth

· Wide range of delivery, meet the delivery needs of most cell types

· High delivery efficiency

· RNP complex as the delivered cargo and significantly reduce the risk of off-target and genomic integration

· Optimized formulation enables high encapsulation, production can be achieved in laboratory

Fig.2 Knockdown efficiency in different cell types （Hepa1-6 Cell, Primary hLESC, RD Cell, K562 Cell, etc. ）

Applications

·Gene edited cell lines with less off-target effect

·Gene editing in stem cells

·Gene editing in primary cells and other difficult-to-transfect cells

·Gene editing in cells derived from other species(e.g., mouse)

For Research Use Only.