"Phages attack, phages attack Every generation, every generation Survival of the remembering" "Repeats in the sequence, repeats in the sequence Twenty-nine base pairs, twenty-nine base pairs What are these palindromes? What are these palindromes?" "Between the repeats, between the repeats Foreign DNA stored, foreign DNA stored Thirty-two nucleotides each Fragments of the enemy" "Cas-1, Cas-2 cutting Cas-9 loaded, ready Molecular sentries Molecular sentries" "Pre-crRNA transcribed, pre-crRNA transcribed From the CRISPR array, from the CRISPR array" "RNase-III cuts, RNase-III cuts Cas-6 trims precisely Guide RNA matured Twenty nucleotides active" "Scan the genome, scan the genome PAM sequence required NGG motif, NGG motif Protospacer Adjacent Motif" "HNH domain cuts the target RuvC cuts the non-target Double-strand break, double-strand break Cellular repair begins" "What if we could program this? What if we could program this? Custom guide RNAs Any target we choose" "Fuse the tracrRNA, fuse the tracrRNA Single guide RNA sgRNA synthetic, sgRNA synthetic One molecule, two functions" "Change the PAM specificity Change the PAM specificity SpCas9, SaCas9 Cpf1, Base editors" "Twenty nucleotides decide Twenty nucleotides decide Where the scissors cut Where the scissors cut" "Lipofection, electroporation Viral vectors, microinjection Into the nucleus, into the nucleus Cas9 ribonucleoprotein" "Non-homologous end joining NHEJ introduces errors Homology-directed repair HDR preserves precision" "GUIDE-seq reveals the miscuts CIRCLE-seq maps the genome Specificity ratios, specificity ratios On-target versus off" "Sickle cell correction Beta-thalassemia cure Leber amaurosis, Leber amaurosis Light restored to darkness" "Drought-resistant wheat Disease-resistant rice Non-browning mushrooms Allergen-free peanuts" "Knockout mice in weeks Knockout mice in weeks Not years of breeding Cellular models of disease" "Adenine to guanine Cytosine to thymine Prime editing inserts Deletions without breaks" "dCas9 catalytically dead dCas9 catalytically dead fused to methylases Chromatin remodeling" Final resolution - all disciplines and time scales present: [All voices: chorus] "From archaea hot springs To human clinical trials Three-point-eight billion years To twenty-five years of engineering Bacterial memory becomes Human molecular scissors Ancient immunity becomes Precision medicine The code that protects Becomes the code that heals CRISPR-Cas revolution CRISPR-Cas revolution"
