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Identifying the best method for delivering the CRISPR-Cas9 system into the cells of interest is the first step in the gene editing workflow. When considering which transfection method to use, transfer efficiency and subsequent cell viability are important factors. Transfection efficiency optimization, construct design, delivery method assessment, and host line selection are some important factors to be considered.
Creating a custom gene-modified cell line starts with evaluating the transfected cell pool. Using the CloneSelect® Imager, effectively screen the edited from the unedited using different selection methods like antibiotic-based, fluorescent protein reporter-based, antibody-tagged cell sorting, and others. The successfully transfected/screened cell pool is then expanded for further monoclonal cell line development.
The DispenCell Single-Cell Dispenser and its dedicated sensing tips can be used to distribute single cells into wells gently and efficiently. After dispensing, the DispenSoft software performs single-cell quality control post-processing cost-effectively.
Documentation of monoclonality (a regulatory metric for therapeutic cell lines) is typically image-based, whereby an image of a single cell is recorded and included in regulatory filings. Many researchers now routinely use imaging systems, such as the CloneSelect Imager, to verify monoclonality at day 0, and monitor cell growth in cell culture media.
Our SpectraMax i3x Multi-Mode Microplate Reader with MiniMax 300 Imaging Cytometer can be used to assess transfection efficiency, monitor cell growth, quantify DNA and protein, and validate CRISPR/Cas9 edits through ScanLater Western Blot analysis. High-quality images of autophagosomes can be acquired using the ImageXpress Micro Confocal System while the MetaXpress HCI software can identify and quantitate individual autophagosomes from every cell, allowing us to analyze phenotypic changes occurring from the CRISPR/Cas9 gene edits.
