Lenti-ONE Cas9

APPLICATIONS

  • Angiogenesis
  • Apoptosis, Oxidative Stress
  • CAR T-cells
  • CRISPR
  • Differentiation
  • Dominant Negative Mutants
  • Neurology, Ophthalmology
  • Optogenetics
  • Recombinases, Editors
  • Reporters and selection
  • RNA Interference

TRANSGENES

Lenti-ONE Cas9

Lenti-ONE GFP carries the transgene green fluorescent protein. Its expression can be assessed by fluorescent microscopy or by flow cytometry, thus giving valuable informations about the transduction efficiency of the vector.

Lenti-ONE Cas9

DESCRIPTION

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.

Cas9

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.

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