We are looking for challenges to test our superfast FluidFM cell editing technology (https://www.cytosurge.com/applications/crispr-cell-line-development) to generate CRISPR-edited monoclonal cell lines. Stop wasting your time to generate your cellular models but use them to advance your research projects! By directly injecting RNP complexes to the nucleus of single cells, we can edit in principle any adherent cell line with higher efficiency, speed and no detectable off target editing!
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Certain genes in Plasmodium falciparum are notoriously difficult to modify by most editing approaches. And more often than not, the efficiency is poor. One of the first attempts for gene editing by delivery of complexed Cas9-gRNA ribonucleoprotein in Plasmodium was to create a point mutation in pfatp4 gene. Although the authors got the mutant, it was against a background of false positive clones. The only way to enrich the edited population of parasites was to use an pfatp4 inhibitor that was insensitive to the mutated Pfatp4 protein. This suggests that although genome edits are possible, the efficiency of the technique is poor. Therefore, it would be really useful to improve this tool for use in Plasmodium.
Ref: Crawford ED, Quan J, Horst JA, et al. Plasmid-free CRISPR/Cas9 genome editing in Plasmodium falciparum confirms mutations conferring resistance to the dihydroisoquinolone clinical candidate SJ733. PLoS One 2017;12:e0178163.
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