![]() In this chapter, various CRISPR/Cas-based approaches are discussed with emphasis on CRISPR/Cas9 vector platforms, multiplex editing strategies, analysis methods for induced mutations, and its applications in medicinal plants. This methodology is based upon the type II adaptive immunity response of prokaryotes, which comprises of a CRISPR-associated (Cas)9 protein and an engineered sgRNA that specifically targets the nucleic acid sequence to induce selective mutagenesis. Due to its wide application in gene mutagenesis, transcriptional regulation, high efficiency and easy manipulation, several plant-specific CRISPR/Cas9 vector systems have been designed. CRISPR/Cas-based genome editing is an emerging technique that utilizes artificially engineered nucleases for digesting DNA at targeted locations in the genome for high-throughput biotechnology-based breeding of valuable medicinal plants. The sequence-specific nucleases of TALENs, ZFNs, and Cas are advance genome editing tools which produce user-defined plants. However, recent biotechnology-based breeding techniques, i.e., genome editing diversify the platform to develop custom-designed medicinal plants. At present, traditional biotechnology techniques have usually been exploited for medicinal plant breeding.
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