CRISPR is a gene-editing method used for rapid diagnostics and precise delivery of genetic material into cells.CRISPR (Clustered Regularly Interspaced Short Palindromic Repeats) is a bacterial immune system turned into a universal tool for genetic engineering. It consists of two components: a guide RNA (sgRNA) complementary to the target DNA segment, and the Cas endonuclease (most often Cas9). The complex scans DNA, recognizes the target site by complementarity, and creates a double-strand break. Repair of the break via non-homologous end joining leads to insertions/deletions, disabling the gene; homologous recombination with a donor template allows precise changes to be made. This technology is significantly simpler and cheaper than previous methods.
The idea of genome editing with nucleases evolved from zinc fingers and TALEN, but the discovery of CRISPR was a revolution. Francisco Mojica in 1993 discovered repetitive sequences in the archaeal genome, later their function in immunity was elucidated. In 2012, Jennifer Doudna and Emmanuelle Charpentier reprogrammed the system for laboratory use. Feng Zhang and colleagues in 2013 first applied CRISPR for editing eukaryotic genomes. For this, Doudna and Charpentier received the Nobel Prize in Chemistry in 2020.
The mechanism is based on complementary binding of RNA to DNA and subsequent hydrolysis of phosphodiester bonds. Cas9 contains two nuclease domains: HNH cleaves the strand complementary to the RNA, and RuvC cleaves the non-complementary strand. The guide RNA consists of crRNA (containing a spacer complementary to the target) and tracrRNA, necessary for binding to Cas. Combining them into a single sgRNA simplified use. The DNA break triggers a cellular response: NHEJ quickly ligates the ends, introducing insertions/deletions, which often inactivates the gene; HDR requires the presence of donor DNA and occurs less frequently, but allows precise editing. Specificity is limited by the thermodynamic stability of the hybrid and the possibility of mismatched binding, but engineered Cas mutations (HiFi-Cas9) reduce off-target cleavage.
CRISPR technology is used to treat hereditary diseases, create pest-resistant plants, and modify animals for research.
With CRISPR, pigs resistant to African swine fever have already been created, and human embryo genes have been edited to correct a mutation causing heart disease.



















