CRISPR Knock-in Through Direct Nuclear Delivery

Current knock-in capabilities support small insertions up to approximately 120 base pairs using ssODN donors with 60 nucleotide homology arms. This enables endogenous epitope tags like HA or FLAG, precise point mutations, and silent PAM-disrupting substitutions.

FluidFM delivers CRISPR RNP and donor templates directly into the nucleus, bypassing cytoplasmic degradation and ensuring immediate co-localization at the break site. Unlike electroporation's stochastic uptake, FluidFM provides metered dosing with consistent nuclear concentrations, reducing cytotoxicity while maximizing HDR engagement

Yes, the FluidFM-based CellEDIT service specializes in hard-to-transfect cells. The direct intra-nuclear delivery method overcomes traditional transfection barriers, making knock-in possible in previously challenging cell types.

Standard QC includes allele-specific PCR assays and Sanger sequencing to verify precise integration, viability assessment, and mycoplasma testing. Matched control clones undergo identical injection without editing. Optional off-target evaluation and monoclonality documentation are available on request.

A knock-in cell line has a specific gene or DNA sequence inserted at a targeted genomic location, while knockout removes or disrupts a gene. Knock-in allows controlled expression of new or modified genes at physiological levels.

Yes, multiplex editing capabilities allow combining single-gene knockouts with knock-ins when feasible. This enables disrupting negative regulators while installing reporter tags or creating null backgrounds with specific rescue mutations. Feasibility is case-dependent and assessed during initial consultation

FluidFM technology enables direct intra-nuclear delivery of CRISPR reagents, bypassing cellular barriers and delivering components directly to the nucleus. This approach enhances efficiency and reduces off-target effects, particularly in hard-to-transfect cells.

The CellEDIT service includes gRNA design, edited and wild-type clones, sequencing validation, and quality control. Customers receive custom monoclonal cell lines with validated insertions.

Knock-in cell lines are used for disease modeling, drug discovery, gene therapy research, synthetic biology applications, neuroscience studies, and agricultural biotechnology. They enable precise study of gene function under physiological conditions.

Yes, FluidFM intranuclear injection is particularly suitable for delicate and hard-to-transfect cell lines including primary cells and iPSCs. The gentle, vector-free delivery preserves viability where conventional electroporation or lipofection methods fail or cause excessive cytotoxicity.

We specialize in precise small knock-ins (≤ ~120 bp) like epitope tags (HA, FLAG) and precise point mutations using ssODN donors via HDR.

FluidFM injects RNPs and donors directly into the nucleus, ensuring immediate co-localization and bypassing cytoplasmic degradation for higher rates.

No, Cytosurge does not claim any IP rights on your cells or specific edits. You retain full ownership of your engineered cell lines.