Lenti-ONE Cas9 contains the cDNA encoding for protein-9 nuclease from Streptococcus pyogenes. (Sequence, WP_010922251.1, 4128bp). Coexpression of Cas9 with an artificial single guide RNA (sgRNA) which fuses the crRNA with the tracrRNA offers the possibility to target and modify DNA sequences of interest.
CRISPR-Cas9 genome editing technology
Lenti-ONE vectors provide an efficient method to drive the suitable level of Cas9 expression. Lenti-ONE Cas9 is available at very high titer >1E9TU/mL, with several packaging options. Thanks to multiple vector configurations, you can build it with a wide range of options, such as:
- Various ubiquitous or specific promoters to adapt the expression level and target particular cell types
- ON/OFF switch to control its expression
- Polycistronic expression cassette to express several gRNA and/or protein of interest
- ANCH/OR system to live track your Lenti-ONE in living cells
- Types of vectors (integrating, non-integrating or RNA) to induce stable or transient expression
Examples of Applications
Lenti-ONE Cas9 can be used alone or co-transduced with other Lenti-ONE vectors.
- Alone, integrating Lenti-ONE Cas9 can transduce cells to produce a cell line that will stably express Cas9. This cell line can then be used to try multiple gRNAs.
- Co-transduced with Lenti-ONE gRNA, it will induce genome editing thanks to Non-Homologous End Joining (NHEJ) and knock-out the target gene
- Insertion of a sequence of interest is also a posibility via Homology Directed Repair (HDR) pathway. A DNA repair template is then necessary. This template contains flanking regions homologous to the target locus. It can be brought with a Lenti-ONE using a tri-transduction with three different vectors, each one bringing Cas9, gRNA and the template DNA. Bi-transduction is also possible if Cas9 and gRNA are brought with an all-in-one vector Lenti-ONE Cas9-gRNA, and the DNA template with a second vector.