CRISPR Cas9 Gene Editing with FluidFM®

In this article, explore the capabilities of the FluidFM technology for CRISPR Cas9 gene editing.

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A vision of CRISPR gene editing with FluidFM 

To access the nucleus and alter the genetic code, CRISPR complexes require an effective delivery method.

CRISPR-Cas9 Gene Editing with FluidFM®
Practically speaking, CRISPR genome editing represents the actions of accurately cutting DNA and allowing natural repair mechanisms to occur. [1] These actions enable the addition, the removal of genetic information or even the alteration at a specific location within the genome. The ease with which CRISPR-Cas9 can be tuned to recognize new genomic sequences has driven a revolution in genome editing that has accelerated scientific breakthroughs and discoveries in disciplines as diverse as synthetic biology, human gene therapy, disease modeling, drug discovery, neuroscience, and the agricultural sciences. [2] 

In the literature, multiple gene editing techniques exist. All those methods present a common challenge: find the right cell transfection method to access the nucleus and alter the genome while keeping the cell alive. Despite being the best approach for research areas where multiple simultaneous edits are required within the same cell line or organism, CRISPR gene editing still encounters limitations due to low HDR efficiency and high off target.  Nowadays, the FluidFM technology offers an unique in vitro solution to improve the efficiency and applicability of CRISPR across a variety of cell types and for cell line development.

CRISPR-Cas9 Gene Editing with FluidFM®

Why choosing FluidFM® for CRISPR Gene Editing?

Direct intra-nuclear Injection

Reach an unprecedented precision delivery and a high CRISPR HDR efficiency due to a ultra-gentle transfection method based on nano-injection.

High Cell Viability

The gentle injection or extraction ensure your cell viability. Especially suitable for hard-to-transfect cells and rare cell types.  

Continuous Cell Monitoring

Easily introduce multiple gene edits in one go while maintaining your cells alive. Fasten your stable monoclonal cell lines generation!

Applications of CRISPR genome editing with FluidFM®

Case study: Multiplex CRISPR editing and monoclonal cell line development

FluidFM nano-injection overcomes delivery limitations of current CRISPR gene editing methods, accelerates cell line development cycles, and is poised to significantly broaden multiplexing capabilities. Pharmaceutical and biological research as well as biologics manufacturing rely on genetically modified cell lines with genes that have been modified to induce the desired phenotype. With the discovery and development of gene editing technologies like CRISPR, the potential of doing multi-loci edits has received much interest but has proven to be a tedious and long process. In the following, we demonstrate the generation of a monoclonal multiple Knock-Out cell line in less than three weeks with the help of the FluidFM technology. 

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Conclusions - Take Away Message

CRISPR Gene Editing

Currently, the most versatile gene editing method available on the market.

The FluidFM Technology

The only single-cell transfection method that provides direct single-cell intra-nuclear injection, extraction or manipulation with a high HDR efficiency  

Versatile Cell Engineering

Full control gene editing OR reliable and reproducible monoclonal cell lines? Everything is possible with the FluidFM Technology!

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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.