CRISPR/Cas9 Lentivirus Production Service

A significant recent advance in genome engineering is the development of the CRISPR/Cas9 system for nuclease-based genome editing and also for transcriptional regulation. In this system, a chimeric single guide RNA (sgRNA) is utilized to direct the Cas9 protein to predefined DNA sequences. The sgRNA can be targeted to any desired DNA sequence by exchanging the 20 bp protospacer that confers targeting specificity through complementary base pairing with the desired DNA target. The CRISPR/Cas9 system can simultaneously target multiple distinct genomic loci by co-expressing a single Cas9 protein with two or more sgRNAs, making this system uniquely suited for multiplex gene editing or synergistic activation of target genes.

Creative Biogene has created the QVirus™ Platform to rapidly generate lentiviral Cas9 constructs for multiplex gene manipulation. This platform readily enabled multiplex gene editing and transient or sustained transcriptional activation in human cells. The lentiviral CRISPR/Cas9 system is useful for both basic science and therapeutic applications by enabling highly efficient and tunable genomic manipulations in cell types that are difficult to transfect, such as primary cells and progenitor cells. Additionally, the vector will be useful for using multiplex gene editing for modeling complex diseases caused by combinations of mutations.

Achieve permanent 100% knockout in your cell line by using QVirus™ Platform´s lentiviral-based CRISPR constructs. Expression of the sgRNA and Cas9 are stable and can be used in dividing or non-dividing cells or whole model organisms. We are offering a Single-Vector as well as a Two-Vector CRISPR-Cas9 System service:

Single-Vector CRISPR-Cas9 System

  • Co-transduction not necessary as sgRNA and Cas9 nuclease are expressed in one vector
  • Lentiviral vectors contain antibiotic selection to ensure stability
  • Lentiviral vectors integrate into the host cell genome and are passed onto daughter cells
  • Cells can be treated, grown for many passages, frozen and thawed - the lentiviral construct remains in the host cells

Two-Vector CRISPR-Cas9 System

  • Pre-transduce with Cas9 lentivirus to decrease noise in your knockout screen
  • Tet-inducible and constitutive U6 expression available
  • Product high-titer sgRNA lentivirus for efficient transduction
  • Vectors have different antibiotic resistance genes to ensure stability and easy selection

Advantages

  • High efficient Cas9 expression delivery with markers
  • No need for tedious cloning work or vector construction
  • Best nuclear penetrating for Cas9 enzyme
  • Allow multiple gene editing at the same time

QVirus™ Platform can produce high yields of lentiviruses encoding the components necessary for CRISPR/Cas9-mediated genome editing (i.e., sgRNA and Cas9 nuclease) for delivery to mammalian cells that are difficult to transfect. If you have any special requirements, please feel free to contact us.

Reference
1. Kabadi A M, et al. Multiplex CRISPR/Cas9-based genome engineering from a single lentiviral vector. Nucleic Acids Research, 2014, 42(19):e147-e147.

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