FluidFM® | CRISPR Cell Line Engineering
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Overcome Cell Line Engineering Delivery Limitations with FluidFM
For multiplex genome editing and fast cell line development.
The discovery of CRISPR-Cas9 for targeted gene engineering has revolutionized life sciences. Regardless of its broad introduction across many disciplines in biology, several challenges persist in editing cell lines, particularly for sensitive cells, or when multiple edits are required.
FluidFM offers a unique in-vitro gene editing supporting tool to improve the efficiency and applicability of CRISPR across a variety of cell types and for cell line development.
Gentle
Ultra-gentle transfection method based on nano-injection
High efficiency
High CRISPR HDR efficiency due to direct intra-nuclear delivery
For sensitive cells
Especially suitable for hard-to-transfect and rare cell types
Multiplexing
Easily introduce multiple gene edits in one go
Fast cell line development
Generate stable monoclonal cell lines
How FluidFM accelerates your CRISPR Cell Line Engineering
No need for carriers or vectors, no limitations by template size
As the CRISPR reagents are directly injected into the nucleus, there is no need to spend time and costs on designing complex plasmids. Also, when working with large repair templates or larger nucleases, there is no size limit as with viral carriers.
Neuron expressing GFP 24 h after injection of a plasmid encoding GFP using FluidFM. Courtesy of Sen Yan, Jinan University, Guangzhou, China.
Especially suited for hard-to-transfect cells
The direct intra-nuclear injection with FluidFM is a very gentle and efficient transfection method. As the insertion of a FluidFM Probe does not compromise cell viability, it can even be used for injecting plasmids, gRNAs or CRISPR-complexes directly into the nucleus of many hard-to-transfect cells with exceptional viability, including stem cells, primary cells and neurons.
Straight forward multiplexing & stack editing
The FluidFM Nanosyringe can be loaded with and deliver hundreds of different gRNA simultaneously and directly into the nucleus of a cell, for highly efficient multiplexing. However, if you prefer stack editing instead, the gentle injection procedure also enables consecutive editing of the same cell.
4 serial injections into the same cell. Lucifer Yellow was used to calculate the volume of solution injected.
Measured volumes of fluorescent CRISPR-Cas9 complexes after injection into mouse primary hepatocytes.
Maximizing HDR efficiency while minimizing off-target & side effects
Direct co-injection ensures that all CRISPR components are delivered simultaneously and at the right concentration into the nucleus. In addition, FluidFM enables quantification of the injected volume, allowing you to calculate exactly how many copies were delivered. This quantification is ideal for evaluation of dose/response relationships to effectively maximize HDR efficiency while minimizing off target effects.
Monoclonal cell line development within 3 weeks
With the FluidFM single-cell approach, you already start from a single cell clone. The FluidFM cell line development workflow, which combines gentle and accurate single cell isolation with the high efficiency of CRISPR gene editing, provides you with a stable, monoclonal cell line within 3 weeks, compared to 10-14 weeks with conventional approaches. Save your precious time, use FluidFM.
Save resources, time & money
CRISPR gene editing with the FluidFM workflow is done on a single cell level. The FluidFM nanosyringe is loaded with only 1µL, and only a dozen of cells is needed per experiment. As such, only a very low quantity of expensive reagents and precious cells are required to obtain your stable, monoclonal cell line by in less than three weeks starting from the day of transfection until the clones are characterized. Start with the needle, not the haystack.
CRISPR Cell Line Engineering with FluidFM - How it works.
The core principle of CRISPR editing and monoclonal cell line development with the FluidFM technology are the hollow force-controlled probes. The variety of available FluidFM Probe tips and aperture sizes enables cell line development less than three weeks:
The FluidFM OMNIUM
The FluidFM OMNIUM is a highly automated, easy to use system for single cell research such as CRISPR genome engineering and monoclonal cell line development.
Stand-alone system
Stand-alone system with temperature and CO2-controlled incubator and extensive automation.
Intuitive software
Software with dedicated workflows, automation, and point-and-click cell selection.
Day 1: FluidFM injection
Select the nuclei of a few cells through point and click. The system will automatically inject the selected cells at a rate of 5 cells per minute. Optionally, a fluorescent marker can be co-injected to facilitate monitoring of the best candidates.
Day 2: FluidFM pick & place
24 hours after injection, analyze the injected cells using the saved coordinates by the software and select the best candidates for isolation. The system will pick each cell and place it into a separate well.
As of day 3: Culture expansion
Culture the picked, monoclonal cells into a colony. After expansion, the monoclonal colonies can be collected and analyzed for the targeted sequence with Sanger sequencing.