Life Science AllianceGenomicsPeer-reviewed journal article
Fast but not furious: a streamlined selection method for genome edited cells
Haribaskar RamachandraniD, Soraia MartinsiD, Zacharias KontarakisiD, Jean KrutmanniD, Andrea RossiiD
Abstract
In the last decade, Transcription Activator-Like Effector Nucleases (TALEN) and Clustered Regularly Interspaced Short Palindromic Repeats (CRISPR) based genome engineering have revolutionized our approach to biology. Due to their high efficiency and ease of use, the development of custom knock-out and knock-in animal or cell models is now within reach for almost every laboratory. Nonetheless, the generation of genetically modified cells often requires a selection step, usually achieved by antibiotics or fluorescent markers. The choice of the selection marker is based on the available laboratory resources, such as cell types, and parameters like time and cost should also be taken into consideration. Here, we present a new and fast strategy called MAGECS (magnetic-activated genome edited cell sorting), to select genetically modified cells based on the ability to magnetically sort surface antigens (i.e. tCD19) present in Cas9 positive cells. By using MAGECS, we successfully generated and isolated genetically modified human induced pluripotent stem cells (hiPSCs), primary human fibroblasts, SH-SY5Y neuroblast-like cells, HaCaT and HEK293T cells. Our strategy expands the genome editing toolbox by offering a fast, cheap, and an easy to use alternative to the available selection methods.
- Citations
- 45
- FWCI
- 2.51
- Field percentile
- top 8%
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