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  Indian J Med Microbiol
 

Figure 1: Illustration showing different steps in Clustered Regularly Interspaced Short Palindromic Repeat-CRISPR-associated protein 9 (CRISPR–Cas9)-mediated gene editing. Cas9 and two single-guide ribonucleic acids (sgRNAs, blue and maroon) were transfected into target cells. Inside cells, complex of sgRNA and Cas9 bind to target sequences. The sgRNA–DNA complex is cleaved by endonuclease activity of Cas9 generating double-strand breaks (DSBs). For homology direct repair (HDR, left), a donor template containing normal deoxyribonucleic acid (DNA) region is also transfected, which provides homologous flanking DNA to repair the cleavage. In nonhomologous end joining (NHEJ, right), cellular repair machinery tries to repair DSB leading to a mixture of DNA sequences. Protospacer adjacent motif (PAM) sites are not mentioned to simplify details. DSB = double-strand break, HDR = homology direct repair, NHEJ = nonhomologous end joining, DNA = deoxyribonucleic acid, sgRNA-1 = single-guide ribonucleic acid-1, sgRNA-2 = single-guide ribonucleic acid-2, Cas9 = CRISPR-associated protein 9

Figure 1: Illustration showing different steps in Clustered Regularly Interspaced Short Palindromic Repeat-CRISPR-associated protein 9 (CRISPR–Cas9)-mediated gene editing. Cas9 and two single-guide ribonucleic acids (sgRNAs, blue and maroon) were transfected into target cells. Inside cells, complex of sgRNA and Cas9 bind to target sequences. The sgRNA–DNA complex is cleaved by endonuclease activity of Cas9 generating double-strand breaks (DSBs). For homology direct repair (HDR, left), a donor template containing normal deoxyribonucleic acid (DNA) region is also transfected, which provides homologous flanking DNA to repair the cleavage. In nonhomologous end joining (NHEJ, right), cellular repair machinery tries to repair DSB leading to a mixture of DNA sequences. Protospacer adjacent motif (PAM) sites are not mentioned to simplify details. DSB = double-strand break, HDR = homology direct repair, NHEJ = nonhomologous end joining, DNA = deoxyribonucleic acid, sgRNA-1 = single-guide ribonucleic acid-1, sgRNA-2 = single-guide ribonucleic acid-2, Cas9 = CRISPR-associated protein 9