CellEDIT - CRISPR Knockout Cell Line Service
Custom CRISPR Knockout Cell Line Service through direct intra-nuclear delivery.
Let us support you with high-quality knockout gene edits, starting from a single cell.
Up to now, CRISPR gene editing is the fastest and most direct approach to achieve the knockout of a specific gene – the modification of the DNA to stop irreversibly the expression of a gene. Now, combining CRISPR gene editing with a vector-free cell transfection, that can deliver the CRISPR complexes directly in the nucleus of a cell, brings a new paradigm in genome engineering.
The CellEDIT workflow, acclaimed for its gentle direct intranuclear CRISPR-Cas9 delivery technique, provides a new version of precision gene editing. This accurate approach minimizes cellular impact and offers unrivaled control over genetic modifications, enabling the creation of stable and high-quality monoclonal mammalian cell lines.
Such high-quality is obtained thanks to the injection of a controlled volume of CRISPR-Cas9 RNP (Ribonucleoprotein) directly into the nucleus, resulting in a reduced residence time until subsequent degradation. Consequently, the fine handling of the RNP volume injection allows to reduce off-targeting effect.
At CellEDIT, we provide you with knockout monoclonal cell lines suited to your project needs in 10 weeks.
Vector-free editing by intra-nuclear delivery
Minimized Off-Targets
Efficient on Hard-to-transfect cells
Knockout Engineered Cell Lines
With CellEDIT, gain access to high-quality CRISPR Knockout cell lines in 10 weeks. The CellEDIT workflow streamlines the efficient engineering and development of knockout monoclonal cell lines via direct intra-nuclear injection, making it ideal for immortalized, cancer, and hard-to-transfect cells.
Immortalized Cell Lines
Learn how the CellEDIT workflow was used to generate 5 monoclonal Hprt knockouts in C2C12 cell line through direct intra-nuclear injection of only 51 cells.
Cancer Cell Lines
Discover how the CellEDIT workflow was used to produce 3 monoclonal HPRT1 knockouts in SK-MES-1 cell, a notoriously hard-to-transfect cell line.
Multiplex Gene Editing
Find out how the CellEDIT workflow boosted the editing efficiency while preserving cell characteristics and functionality in single and multiple KO CHO-K1 clones.
Applications
Gene knockout is a critical tool in biological research, allowing scientists to understand the specific role and function of individual genes by selectively disabling them. This method provides insights into disease mechanisms, contributes to drug development, and reveals complex gene interactions and regulatory networks, enhancing our understanding of biological systems. As an example, here are some specific ways gene knockout is used in in-vitro disease modeling:
Disease Mechanisms
By knocking out a gene suspected to cause a disease, researchers can create cell models that mimic the disease state. This allows them to study the cellular and molecular changes that result from the loss of that gene's function, leading to a better understanding of how the disease develops and progresses.
Gene Function
Knocking out a gene helps determine its function. If a gene is suspected to be involved in a particular cellular process, knocking it out can confirm whether the process is affected by the absence of that gene.
Target Identification
Gene knockout can help identify potential therapeutic targets. If knocking out a gene improves disease symptoms in a model, it suggests that inhibiting the function of that gene in humans might also be beneficial.
Drug Validation
If a drug targets a particular gene product, knocking out that gene can help validate whether the drug is acting specifically on its intended target. If the drug is still effective in the absence of the target gene, it might indicate that it works through off-target effects.
Drug Efficacy & Toxicity
Using disease models with knocked-out genes, researchers can also assess the potential efficacy and toxicity of experimental treatments.
Yet, performing gene knock out remains a challenge for many researchers, leading to the development of gene knock out cell line services.
Why choosing a service to perform your engineered knockout cell line?
Utilizing a knockout gene editing service provides a researcher with access to expert knowledge and sophisticated techniques, ensuring precise, cost-effective, and successful gene manipulation.
The CellEDIT Cell Line Engineering Service, with its know-how and continued support throughout your project, provides accuracy, reliability, and additional reproducibility to your biological research by providing you with high-quality knockout edited mammalian cell lines in 10 weeks.
The success of a CRISPR knockout gene editing approach relies on several parameters, including the way CRISPR complexes are introduced in the nucleus of the cell, that is the cell transfection.
The CellEDIT workflow revolutionizes cell transfection by adopting a vector-free approach. Unlike conventional methods that rely on stochastic delivery to the entire cell population, our technique offers a precise and targeted solution for high-quality cell line generation by injecting CRISPR complexes directly into the nucleus, thanks to our unique patented FluidFM technology.
This breakthrough approach employs the FluidFM Nanosyringe, enabling a gentle and controlled nano-injection of CRISPR Cas9 RNP directly into the nucleus. Utilizing Cytosurge's patented FluidFM technology (Fluidic Force Microscopy), the process integrates five main steps: single-cell plating, single-cell injection, monoclonal expansion, validation of edits, and finally, cryopreservation. This approach, characterized by using microscopic channels within force-sensitive probes, is adaptable to various knockout cell line generations, encompassing both easy-to-handle and hard-to-transfect mammalian cells.
By bypassing the limitations of other delivery methods, we maximize the efficiency of knockout gene editing by injecting accurately a set volume of RNP into the nucleus, reducing off-targeting effect. The precise and controlled targeting of the nucleus empowers researchers with unparalleled control and accuracy in their experiments. Ultimately, this comprehensive workflow paves the way for the creation of stable monoclonal cell lines, enabling gene knockout and multiplexing.
Resources
Educational
Technical
Introduction to CRISPR Knockout Gene Editing with CellEDIT
An Overview of CellEDIT' CRISPR Cell Line Development Services
CellEDIT' CRISPR Cell Line Development Workflow
CellEDIT's Engineered Cell Lines
CRISPR-Engineered U2OS Cell Line
CRISPR-Engineered MDA MB 231 Cell Line
CRISPR-Engineered A549 Cell Line
CRISPR-Edited HEK293 Cell Line
CRISPR-Engineered C2C12 Cell Line
Media & Downloads
On-Demand CellEDIT's CRISPR Cell Line Engineering Webinar
CellEDIT's Application Note n*1 - Introducing the CellEDIT Workflow
CellEDIT's Application Note n*2 - Overcoming the hard-to-transfect cell line hurdle
Case Study - Streamlined U2OS Cell Line Modification with the CellEDIT Service Workflow, featuring our CellEDIT Customer: Dr. Kanstantsin Siniuk.
Open Access Publication featuring CellEDIT: Antony, Justin S., Anabel Migenda Herranz, Tahereh Mohammadian Gol, Susanne Mailand, Paul Monnier, Jennifer Rottenberger, Alicia Roig‐Merino et al. " Accelerated generation of gene-engineered monoclonal CHO cell lines using FluidFM nanoinjection and CRISPR/Cas9" Biotechnology Journal 19, no. 4 (2024): 2300505.