Therapeutic Genome Editing

This volume offers a practical, methods-first guide to therapeutic genome editing across 25 chapters, grouped into seven parts. It opens with the foundational platforms — CRISPR, zinc-finger nucleases, and TALENs — then works through the newer toolkit: Cas12a, anti-CRISPR proteins, base and prime editing, dual-pegRNA strategies, bridge recombinases, Cas transposases, and epigenome editing. From there, the focus shifts to disease. Chapters walk through applications in human iPSC models, corneal and retinal disorders, targeted editing of Genome Wide Association (GWAS) variants, and therapeutic correction of a monogenetic disorder (RPGRIP). Because safety is where translation succeeds or fails, several chapters are devoted to it — assessing genome-wide off-target activity and structural variants with GUIDE-seq, SITE-seq, BLESS, Digenome-seq, and CIRCLE-seq, alongside Bionano optical genome mapping and PacBio sequencing. The volume also reaches into emerging methods: saturation genome editing in organoids, ticker-tape for lineage tracing, minicircle DNA delivery, and GMP manufacturing of AAV vectors for Cas9 delivery. As with every title in the Methods in Molecular Biology series, each chapter pairs essential background with detailed materials, step-by-step protocols, and troubleshooting notes. Therapeutic Genome Editing: Methods and Protocols is written for researchers, trainees, and clinicians moving genome editing from the bench toward the clinic.