I have experiences in RNAi and Cas9 knockout experiments in mammalian cells, but am now facing a little challenge. I have primary chicken and duck endothelial cells that require knockdowns of genes - knockouts of these genes are impossible due to their importance for these cells. I have seen 1-2 papers looking at CMV/Ef1a...
I trying to generate a KO model in ha hTERT line of fibroblast using lentiCRISPR V2 with 3 gRNA. The gRNAs are desining to generate a delection between exons 1 and 3 of my gene. I already performance puromicine selection and try to validate expresion levels of mRNA for me gene in policlonal cell line but I don't get any cha...
In a recent experiment I used lentiviral transduction of CRISPR to knock-out/down a gene of interest in a cell line. As a negative control I used the native lentiCRISPRv2 plasmid (with the 2kb filler) instead of the lentiCRISPRv2 plasmid cloned with a sgRNA against GFP for example. When I added the Puromycin to the cells, m...
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