Gene Editing (CRISPR/Cas9)

Accelerating gene edited cell lines

Learn how the all new CloneSelect® Imager FL can aid in easy detection of successfully transfected cells, cutting cell line development timelines and scaling up your research faster. Reject low transfection efficiency pools at an early stage, confirm and track various CRISPR edits with multi-channel fluorescence detection, and screen cells with accuracy and confidence while reducing the risk of over-passing disturbances with robotics redesign.

Read application note  

Clone productivity screening and titer

An important component in identifying high-value clones is determining productivity of single cell-derived colonies. Screening for productivity using traditional approaches is laborious and time consuming, generally consisting of a multistep process that involves isolating single cells from limiting dilution followed by assessment of titer using ELISA. The ClonePix 2 system combines phenotype selection, single-cell isolation and productivity screening into a single step, resulting in dramatically shorter screening times and increased number of candidates.

• Rapid Screening and Selection of Stable High Producing Clones
• Automated optimization of IgG production in CHO Cells
• Enhanced development of virus-specific hybridomas

Confident assurance of clonality using calcein AM

Confident assurance of clonality using calcein AM with minimal effect on viability

Here, we demonstrate an optimized workflow using the fluorescence reagent, calcein AM, in conjunction with a fluorescence-capable CloneSelect™ Imager that shows similar viability to label-free conditions while simultaneously providing high assurance of clonality.

Read application note  

CRISPR in cell line development

Targeted gene editing, especially with CRISPR, has revolutionized science and has many applications in various fields, mainly in cell line development, cell and gene therapy.

Typical cell-line development requires the screening of tens of thousands of clones to find the best stable cells that yield high amounts of bioproducts. In this application note, we discuss the automation of a mammalian cell-line development workflow—the automated, integrated screening of transfected cells for edits, monoclonality, and growth assessment.

Read application note  

CRISPR/Cas9 genomic editing experiments

The CRISPR/Cas9 gene editing system is a very popular tool for studying gene function due to its relative ease of use and accuracy. Additionally, the system has enormous potential for treating hereditary diseases. Validation of CRISPR/Cas9 gene editing is necessary to ensure that genes of interest are successfully knocked down or knocked out. Here, we demonstrate how Molecular Devices' family of instruments can be utilized in gene editing experiments by using CRISPR/Cas9 to knockdown autophagy-related protein 5 (ATG5) in HEK293 cells.

View poster  

Drug Discovery & Development

The drug discovery landscape is shifting, with more scientists centering cell line development, disease models, and high-throughput screening methods around physiologically-relevant 3D cell models. The reason for this is clear: Using cellular model systems in research that closely mimic patient disease states or human organs can bring life-saving therapeutics to market – faster.

Learn more  

Monoclonality

Cell line development and assurance of monoclonality are critical steps in the process of generating biopharmaceutical molecules, such as monoclonal antibodies. A cell line can be established following the isolation of a single viable cell robustly expressing the protein of interest. A key milestone in this process is documenting evidence of clonality. Documentation of clonality is typically image-based, whereby an image of a single cell is produced and included in regulatory filings.

Learn more 

Single-Cell Sorting

Cell line development requires the discovery of single cell-derived clones that produce high and consistent levels of the target therapeutic protein. The first step in the process is the isolation of single, viable cells. Limiting dilution is a technique that relies on statistical probability but is time consuming. The DispenCell™ Single-Cell Dispenser enables fast, easy, and gentle isolation of cells, as well as provides instantaneous proof of clonality and traceability post-cell dispensing.

• Brochure: DispenCell Single-Cell Dispenser

Transfection efficiency

Transfection efficiency for a fluorescent reporter gene can be monitored in different ways. One way is to measure fluorescence with a microplate reader. This allows one to assess the overall fluorescence level in each test well, but it does not give the percent of cells transfected. A more informative way to assess transfection efficiency is to analyze the cells using an imaging cytometer, where the number of cells expressing detectable fluorescence can be compared to the total cell number. The imaging cytometer has the added benefit of enabling calculation of cell confluence prior to transfection, so that this information can be used as part of assay development.

Read application note  

Validate CRISPR-Edited Cells using Western Blot

CRISPR gene-editing technology requires careful monitoring of the entire process to ensure accurate results. The SpectraMax i3x Multi-Mode Microplate Reader provides a complete solution for analyzing the results of a CRISPR-editing experiment from initial transfection to confirmation of protein knockdown. With the MiniMax cytometer, researchers can assess transfection efficiency by comparing total unlabeled cell counts to counts of fluorescence expressing transfected cells. The ScanLater Western Blot Detection System enables sensitive detection and quantitative analysis of proteins of interest in control and CRISPR-edited cells.

Read application note