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Fig. 3 | Biological Procedures Online

Fig. 3

From: Review of applications of CRISPR-Cas9 gene-editing technology in cancer research

Fig. 3

Basic steps of CRISPR-Cas9 gene screening: First, plasmids were constructed that expressed the corresponding CRISPR components, they were packaged as viruses, libraries were built, and the cancer cells to be studied were infected. Then, uninfected cancer cells were removed by drug selection (e.g., puromycin), and the screened cancer cells were divided into three groups: the day 0 population, drug-treated population (treatment) and control population (mock-drug control, typically treated with a vehicle such as DMSO). After specific drug treatments, genomic DNA was extracted from the transduced cells, and high-throughput sequencing was performed to sequence the sgRNA coding region that the virus had integrated into the host genome. Finally, bioinformatics analysis was performed to compare the sequencing results of the three groups of cancer cells, and the genes that were affected by specific drugs were preliminarily obtained

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