What is a Gene Editing Tool?
Gene editing tools are techniques and technologies that allow for precise modifications of an organism's DNA. These tools enable researchers to insert, delete, or modify specific genes, facilitating studies in genetics, disease mechanisms, and therapeutic development. Among these tools, CRISPR Cas9 has emerged as the most popular due to its simplicity, versatility, and efficiency. CRISPR Cas9 uses a guide RNA to target a specific DNA sequence, where the Cas9 enzyme creates a double-strand break. This break allows for targeted gene editing, making CRISPR Cas9 a powerful tool in various fields of research.
Challenges in Gene Editing
Despite its revolutionary capabilities, gene editing with CRISPR Cas9 faces several challenges:
Low Efficiency: Traditional delivery methods often fail to deliver CRISPR Cas9 complexes effectively into cells, resulting in low editing efficiency.
Off-Target Effects: Inaccurate delivery can cause the potential activation of immune responses and cytosolic degradation.
Delivery Barriers: Introducing CRISPR Cas9 directly into the nucleus is challenging, especially for hard-to-transfect cells, limiting its application range.
Direct intra-nuclear delivery is critical for achieving precise gene editing outcomes, as it ensures that the CRISPR Cas9 complexes reach their intended target without degradation or loss, minimizing off-target effects and maximizing editing efficiency.
Introducing the FluidFM Technology
FluidFM (Fluidic Force Microscopy) is an innovative technology that integrates microfluidics with Atomic Force Microscopy (AFM) to enable precise manipulation at the single-cell level.
FluidFM uses a hollow cantilever with a nanoscale aperture, allowing for the controlled injection or aspiration of small volumes directly into individual cells. This unique approach provides unparalleled precision in delivering biomolecules, such as CRISPR Cas9 complexes, directly into the nucleus, overcoming the limitations of conventional transfection methods.
A. The FluidFM OMNIUM platform is a single-cell manipulation system designed to streamline the use of FluidFM probes in life-science applications. B. SEM picture of the FluidFM Nanosyringe - The sharp apex of the FluidFM Nanosyringe enables gentle probe insertion into a cell to inject precisely a set amount of RNP complexes directly into the nucleus of a cell.
Combining FluidFM with CRISPR: A Game-Changing Gene Editing Tool
In 2022, Cytosurge launched the CellEDIT service – a CRISPR Cell Line Engineering Service that introduces a disruptive workflow to deliver precise quantities of CRISPR Cas9 Ribonucleoproteins directly into the nucleus. This targeted approach enhances CRISPR Cas9 delivery by providing a direct and controlled method to introduce gene-editing complexes into the nucleus, minimizing the risk of immune response activation and cytosolic degradation, thus significantly maximizing the editing efficiency and precision compared to conventional methods.
Vector-free editing by intra-nuclear delivery
FluidFM's nanoscale probe injects CRISPR Cas9 complexes directly into the nucleus, bypassing cellular barriers.
Efficient on Hard-to-transfect cells
The FluidFM Nanopipette injects CRISPR Cas9 complexes directly into the nucleus, making it ideal for engineering cell lines in hard-to-transfect cells.
Reduced Off-Targeting Effect
With FluidFM, all CRISPR complexes are delivered simultaneously and at the right concentration into the nucleus, effectively reducing off target effects.
By combining FluidFM with CRISPR Cas9, researchers achieve higher efficiency, reduced off-target effects, and improved outcomes, making it a game-changing tool for gene editing.
Applications of FluidFM-Enabled Gene Editing Tool
Functional Genomics: FluidFM facilitates precise gene knockout and knock-in studies, enabling researchers to explore gene function and regulation with greater accuracy.
Disease Modeling: By allowing targeted gene modifications, FluidFM helps create accurate cellular models of genetic diseases, advancing our understanding of disease mechanisms.
Therapeutic Development: FluidFM's precision in delivering CRISPR Cas9 complexes optimizes the development of gene therapies, ensuring effective targeting and reduced side effects.
Synthetic Biology: The technology supports the introduction of synthetic genes or circuits into cells, fostering innovation in bioengineering and synthetic biology.
FluidFM's ability to enhance CRISPR Cas9 gene editing offers a powerful tool for life sciences professionals and biological researchers, paving the way for breakthroughs in genetics, disease research, and therapeutic development.