Genomics and Applied Biology 2024, Vol.15, No.5, 223-234 http://bioscipublisher.com/index.php/gab 227 Figure 2 Molecular and chemical characterization of cannabinoid synthase genes in European Cannabis genotypes (Adapted from Fulvio et al., 2021) 4.4 Technological tools for gene mining Advancements in genomic technologies have significantly enhanced the ability to mine functional genes in Cannabis sativa. Techniques such as transcriptomics, proteomics, and next-generation sequencing have been instrumental in identifying and characterizing genes involved in cannabinoid biosynthesis, disease resistance, and fiber production. For example, the use of RNA sequencing (RNA-seq) has enabled the detailed analysis of gene expression profiles in different cannabis strains, revealing key differences in metabolic pathways (Bakel et al., 2011; Romero et al., 2020). Proteomic approaches have further elucidated the diversity of enzymes involved in cannabinoid synthesis and their regulatory mechanisms (Romero et al., 2020). Additionally, the integration of multi-omics data, including genomics, transcriptomics, and metabolomics, has provided a comprehensive understanding of the complex gene networks in cannabis (Wu et al., 2021). 5 Role of Genomic Tools in Breeding Programs 5.1 Use of genomic selection for trait improvement Genomic selection has become a cornerstone in modern breeding programs, leveraging high-throughput DNA marker genotyping and whole genome sequencing to enhance the selection process. This approach allows breeders to predict the genetic value of plants more accurately and efficiently, thereby accelerating the development of new varieties with desirable traits. The integration of genomic selection with traditional breeding methods has shown significant promise in improving yield, disease resistance, and stress tolerance in various crops (Thomson et al., 2022). 5.2 CRISPR-Cas9 and other gene-editing tools in Cannabis breeding The advent of CRISPR-Cas9 and other gene-editing technologies has revolutionized plant breeding, including Cannabis. CRISPR-Cas9, in particular, offers a precise, efficient, and relatively simple method for targeted genome modifications. This technology enables the deletion of detrimental traits and the addition of beneficial ones, making it a powerful tool for functional genomics and crop improvement (Bortesi and Fischer, 2015; Arora and Narula, 2017; Jaganathan et al., 2018). The versatility of CRISPR-Cas9 extends to generating knockouts, precise modifications, and multiplex genome engineering, which are crucial for developing Cannabis varieties with enhanced traits (Arora and Narula, 2017; Ahmar et al., 2020; Zhang et al., 2021). Additionally, the development of CRISPR ribonucleoproteins (RNPs) has addressed some limitations of plasmid-based systems, further enhancing the efficiency and applicability of this technology in Cannabis breeding (Arora and Narula, 2017).
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