CRISPR & Cell Engineering
Rapid and Gentle Single Cell Isolation for CRISPR Cloning & Cell Engineering
The precision gene editing enabled by the use of CRISPR (clustered regularly interspaced short palindromic repeats) technology has great potential for use in life science and biomedicine. Typically, a CRISPR cell engineering workflow involves the transfection of target cells with plasmid vectors containing both the CRISPR-Cas9 gene editing construct and a fluorescent reporter (this enables successfully transfected cells to be easily identified). The next step involves the isolation of edited cells for single cell cloning and clonal expansion. There are two techniques that have traditionally been used for single cell isolation, both of which have certain drawbacks.
- Limiting dilution - the single cell isolation efficiency using this method is highly variable, which may result in multiple cells per well or empty wells. Add to this the possibility of low transfection or editing efficiency and the isolating of CRISPR clones by this method can become hugely inefficient, costly and almost unworkable.
- Fluorescence-activated cell sorting (FACS) - this method of cell isolation is faster than limiting dilutions resulting in higher throughput, however FACS sorting systems operate at high pressure (up to 70 psi), which can result in cell damage or death. Commonly used cell types for CRISPR cloning, such as induced pluripotent stem (iPS) cells, are particularly sensitive to the high pressure and have low survival rates when sorted by FACS. Another downside to using FACS is that it is not easy to use and is prone to contamination.
Rapid Single Cell Isolation
Bio-Techne's single cell sorters are the simplest and fastest way to identify and isolate target CRISPR-Cas9 gene-edited single cells for your cell engineering workflow. And it's all achieved seamlessly, in one step.
Highly Efficient Single Cell Dispensing
Cells are dispensed 1 cell/well at high efficiency into 96 or 384 well plates. The image [right] shows that Bio-Techne instruments are highly efficient and consistent at dispensing single cells (80-90% with mammalian cells) compared to manual pipetting (~30%).
Gentle Cell Sorting
Our single cell sorters can preserve the viability and integrity of cells for better clonal outgrowth. The low-pressure sorting by our instruments is gentle on your precious cells.
Single Cell Cloning Efficiency
The figure shows a comparison of Bio-Techne’s Single Cell Dispenser with traditional methods of sorting cells for clonal outgrowth. Bio-Techne’s Single Cell Dispenser, was used alongside limiting dilution (LDC) and FACS sorting to sort CHO host cells for a single cell cloning protocol sorting. Results highlight the higher cloning efficiencies obtained with Bio-Techne’s platform compared with LDC and FACS.
Empower CRISPR Cell Engineering with Innovative Single Cell Sorting
Discover how scientists are using Bio-Techne's Single Cell Dispensers to empower single cell cloning to generate CRISPR-edited cell lines. Discover the key role that single cell sorting and isolation can play in your cell engineering workflow for a wide range of application from cell biology, to drug discovery and future cell therapy.
CRISPR-Edited Cell Line Development Workflow Improvements
CRISPR-Edited Cell Line Development Workflow Improvements
In this whitepaper Dr Oscar Perez-Leal of Temple University describes how the Bio-Techne Single Cell Dispenser can streamline and accelerate the development of cell line models generated using CRISPR-Cas9 gene editing.
Webinar: Producing High Quality CRISPR-Cas9 Edited hiPSCs
Webinar: Producing High Quality CRISPR-Cas9 Edited hiPSCs
Learn from Dr Maks Prondzynski about optimizing the growth and differentiation of human induced pluripotent stem cells (hiPSCs) into cardiomyocytes to support enhancing cardiac disease modeling.
Breaking Barriers in Cell Engineering with Single Cell Sorting Innovation
This eBook, produced in collaboration with the team at Drug Target Review focuses on how advances in single cell sorting technology are furthering research in CRISPR gene editing and cell engineering. The team shares insights, looks at recent peer-reviewed publications, and talks one on one with the scientists.
Dr Oscar Perez-Leal, MD, Assistant Professor, Temple University, School of Pharmacy
“Utilizing this machine [the Bio-Techne Single Cell Dispenser] has proven instrumental in streamlining the development process of single-cell clones for the cell lines I am working on. It has greatly facilitated the generation of valuable research tools, contributing to the overall progress and efficiency of my work.”
Dr Po-Lin So, Gladstone Institutes
“Our stem cell core facility was looking for ways to speed up our human induced pluripotent cell cloning to generate clonal cell lines and to analyze our gene editing efficiencies in single cells. Our criteria for an instrument for single cell dispensing was 1) sterility, 2) gentle on cells, 3) ease of use, and 4) reasonable cost….[With Bio-Techne,] We are able to reduce the time and effort for generating clonal CRISPR-engineered iPSC lines significantly. The instrument requires very little maintenance and users do not need to be highly trained with the main requirement being good aseptic technique.”
Customer Success Stories
Genentech
C4 Therapeutics
BioMicro Center, MIT
Cytovance Biologics