CRISPR Knock-in Cell Line Service (Human Primary, T Cells, & Blood Lineage Cell Lines)

We offer efficient CRISPR-based stable knock-in cell line generation for hard-to-transfect cell lines such as blood lineage cells, primary T cells and fibroblasts, and more, using multi-approach designs and highly optimized CRISPR knock-in cell line protocols. ASC has generated 1300+ unique cell line models with a wide variety of modifications in 200+ mammalian cell lines, and we are one of the very few CRISPR cell line generation in hard-to-edit cell lines service providers who knows how to work with these difficult cell lines.

Other features include:

• Homozygous/ heterozygous knock-in clones
• Variety of cell lines: cancer, hard-to-transfect, most mammalian species

Workflow includes:

• Cell line evaluation (culture conditions, single cell clonability, etc.)
• gRNA design, construction, and validation
• Donor DNA construction
• Cell line validation, transfection and optimization
• Screening for single cell clones and clone confirmation
• Cell expansion and cryopreservation

Deliverables:

• Two (2) vials of 2 x 10^5 cells/ vial of cell line with targeted insertion either as stable single cell clones or clonal pools (as determined by milestone 1).
• 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: 3-4 months

Applications:

• Disease modeling for immuno-oncology, pharmacogenomic studies
• Drug discovery and drug efficacy and toxicity screening; drug combination studies
• Antibody validation/ screening
• Deriving diagnostic reference standards and materials
• Recombinant protein production in CHO cells
• Generation of TARGATT™ master cell lines by inserting an attP "docking site" for site-specific gene knock-in