CRISPR Gene Editing & FluidFM: Revolutionizing Genome Engineering
Introduction to CRISPR Gene Editing with FluidFM
To access the nucleus and alter the genetic code, CRISPR complexes require an effective delivery method.
While several gene editing techniques exist, each faces the challenge of efficiently accessing the nucleus to modify the genome without harming the cell. Though CRISPR excels in scenarios needing multiple edits within a single cell line or organism, it sometimes falls short due to low HDR efficiency and off-target effects.
Presently, FluidFM technology enhances CRISPR's efficiency and versatility in various cell types and is pivotal for cell line engineering and development.
Applications
The CellEDIT CRISPR Cell Line Engineering Workflow
As technology and biopharmaceutical research evolve, the importance of cell line development has grown, especially in areas like pathway analysis, target validation, and disease modeling. This growth in novel therapeutics has amplified the need for advanced cell line development techniques. In response, scientists are combining traditional transfection methods with CRISPR/Cas complexes for improved cell line development. While older methods typically use batch transfection techniques like electroporation or lipofection, the CRISPR Gene Editing with FluidFM performed via the CellEDIT service offers an innovative single-cell transfection process. This method administers exact amounts of CRISPR Cas9 Ribonucleoproteins directly into the nucleus, reducing potential immune reactions and cytosolic breakdown. As a result, it offers a more efficient and accurate solution than traditional methods.
Application note: The CellEDIT Workflow Facilitates the Efficient Production of Knockout Clonal Cell Lines by Direct Intranuclear Injection
- The CellEDIT workflow successfully generated 4 monoclonal hprt knockouts in C2C12 cell line by direct intranuclear injection of only 27 cells.
- Genotype analysis revealed that 97% of the expanded clones presented gene editing, with 48% of the clones displaying editing on all alleles.
- Direct intranuclear injection proves to be a highly controllable and gentle alternative transfection method for CRISPR-mediated gene editing.
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References
[1] Jinek, Martin, et al. "A programmable dual-RNA–guided DNA endonuclease in adaptive bacterial immunity." science 337.6096 (2012): 816-821.
[2] Li, Hongyi, et al. "Applications of genome editing technology in the targeted therapy of human diseases: mechanisms, advances and prospects." Signal transduction and targeted therapy 5.1 (2020): 1-23.