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The method leverages the **CRISPR-Cas9** system, which acts like a pair of molecular scissors. The Cas9 enzyme, guided by a synthetic guide RNA (gRNA), can precisely target and cut specific DNA sequences in the genome. When Cas9 cuts the DNA, the cell's natural repair mechanisms kick in. There are two main ways the cell can repair the break: non-homologous end joining (NHEJ) and homology-directed repair (HDR). In CRISPR screens, researchers typically exploit NHEJ, which is an error-prone process that often leads to small insertions or deletions (indels) in the DNA sequence. These indels can disrupt the gene's coding sequence, effectively knocking out the gene's function. By creating a library of gRNAs that target different genes in the genome, researchers can generate a diverse pool of cells, each with a different gene disrupted.
