CRISPR Knockout Cell Line Generation
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A comprehensive custom Lentiviral-based Stable Cell Line Generation Service for genetically engineering sensitive cells, primary cells, stem cells and terminally differentiated cells. Lentivirus-based stable cell line generation offers a cost-effective and efficient alternative for generating stable cell line models for gene overexpression, inducible gene expression, gene knockout and gene knockdown.
- Broad tropism lentiviruses to target most mammalian cell types
- Non-integrating lentiviruses for CRISPR-lenti gene knockout
- Choice of selection markers: antibiotic resistance or GFP
- 3rd generation lentiviral systems for the highest biosafety standards
- Gene editing confirmed by NGS or RT-PCR
Cell Line Modifications:
- Knock-in Cell Line Generation: We offer 3rd generation lentiviral system for stable gene knock-in into multiple cell types from most mammalian species.
- CRISPR Knockout Cell Line Generation: We offer non-integrating lentiviruses (IDLVs) for CRISPR-lenti gene knockout or knock down modifications in hard-to-transfect cell lines.
- Cell line validation
- Generation of lentivirus; CRISPR vector construction and lentivirus packaging (if needed)
- Reagent validation
- Lentivirus transduction
- Clone screening and confirmation
- Cell expansion and cryopreservation
Customer should provide:
- Frozen cells: 2 vials of 10^6 cells/ vial OR live cells: 2 x T25 flasks at 90% confluency
- Detailed information regarding target gene and gene modification
- Two vials of 1-2 clones or two vials of pooled clone.
- Optional: negative control non-target gRNA-virus available
- Milestone update/report; final report with detailed description of each procedure, including targeting vector design, construction and validation, transfection condition, genotyping strategy, and results.
Timeline: 3-6 months
- Drug screening
- Functional assays
- Membrane protein screening
- Long-term expression studies
- Antibody immunization boosting
- Gene therapy research