Boost CRISPR efficiency

A solution to CRISPR’s delivery challenges.

FluidFM technology offers a unique tool for gene editing. The stand-alone system enables to efficiently deliver any kind of soluble compound directly into any adherent or suspension cell. Even when it comes to sensitive and hard-to-transfect cells, or when large cargos have to be delivered, FluidFM nano-injection provides the ideal solution.

Deliver all CRISPR components, or any other cargo, simultaneously and at defined concentration directly into the nucleus whilst maintaining high cell viability. This paves the way to use large templates and to easy multiplex- and stack-editing. The stand-alone system also enables optimization of off-target effects and HDR efficiency as the quantity of injected copies can be calculated and therefore dose/response relationships investigated. The injection process is highly automated and reproducible with an intuitive user interface – perform direct delivery with a simple mouse click.

Odoo - Sample 3 for three columns

Higher delivery & HDR efficiency

All CRISPR components are delivered simultaneously and at the right concentration directly into the nucleus. For any cargo & cell type, maintaining high cell viability and paving the way to large genomic editing.

Odoo - Sample 3 for three columns

Optimize off-target effects

As the quantity of injected copies can be calculated, dose/response relationships can be investigated and thus off-target mutations minimized.

4 serial injections into the same cell. Lucifer Yellow was used to calculate the volume of solution injected.

Easy multiplex & stack editing

A FluidFM nanosyringe can be loaded with 100's of different gRNAs. Biology is the limit.

Inject different cocktails at different time points: >95% of cells survive the injection procedure.

Direct delivery with a simple mouse click.

Automated and reproducible injection process with an intuitive user interface. 

 Use Cases 

1. Injection of multiple gRNAs&Cas9 

A fluorescent marker is co-injected into CHO cells to verify successful delivery.

Use Case #1.1. - Direct intra-nuclear CRISPR-Cas complexes direct delivery with FluidFM
  • Multiple gRNAs are simultaneously delivered

  • Injection process maintains >95% viability

2.  Isolation of the clone candidate

Selected cells are isolated into separate wells.

Use Case #1.2. - Direct intra-nuclear CRISPR-Cas complexes direct delivery with FluidFM
  • Monoclonality proof: visual confirmation of the deposition of a single cell

  • Isolation process maintains >95% viability

3.  Clone expansion and analysis

Monoclonal colonies are expanded and the DNA sequenced. 50% of the colonies show a mutation in all the targeted loci.

Use Case #1.3. - Direct intra-nuclear CRISPR-Cas complexes direct delivery with FluidFM
  • Extreme reduction in complexity and development time of modified cell lines starting from few nano-injected cells

Transient expression of target proteins with mRNA

Human Dermal Fibroblast injected with GFP mRNA. 70% of injected cells expressed GFP 24h after injection.

Transient mRNA transfection helps to give fundamental insights on protein functions at a single cell level, such as protein expression and intercellular communication.

 mRNA transfection using FluidFM nano-injection results in:

  • Higher transfection efficiency

  • Lower cell toxicity

  • Faster expression

Image: Human dermal fibroblast expressing GFP, 24h after injection.

Odoo • Text and Image

Odoo • Image and Text

Successful transfection of primary neurons with plasmids

With our system, even primary neurons can be transfected successfully and highly efficiently. The FluidFM nanosyringe directly delivers the complexes into primary neurons - gently and autonomously.

Conventional delivery methods applied to neurons are often inefficient, expensive and toxic to the neuron.

Image: GFP expression in neurons 24h after nano-injection of plasmids in a mouse primary neuron. Courtesy of Jinan University Guangzhou, China.