Industrial Cannabis sativa (Hemp) is a multipurpose revolutionary crop widely cultivated for its seeds, which are rich in oil, CBD content, proteins, carbohydrates, fibers, as well as vitamins and minerals. Genome editing, CRISPR/Cas9 acts as highly precise "genetic scissors" that accelerate traditional plant and animal breeding by modifying specific DNA sequences without introducing foreign (transgenic) DNA. The world's first genome-edited rice varieties are DRR Dhan 100 (Kamala) and Pusa DST Rice 1 developed by the Indian Council of Agricultural Research (ICAR),New Delhi, India. Genome editing, CRISPR/Cas9 revolutionizes Cannabis sativa (hemp) breeding by enabling precise, targeted modifications in a crop traditionally hampered by dioecy and high heterozygosity. CRISPR/Cas9 technology allows for sequence specific editing of the target genome, thereby allowing for precise control over gene modifications and associated traits, in a low cost and straightforward manner. Gene editing has been successfully used to generate climate-resilient crops for various climatic conditions. However, several limitations are descending its overall potential remains limited. The biological functions of most candidate genes are poorly characterized. For practical crop improvement in Cannabis sativa, functional validation serves as a bridge between genomic discovery and breeding applications. The Cannabis sativa genome has long been challenging to genetically manipulate due to recalcitrance in tissue culture and transformation, limiting functional validation of genes and targeted trait improvement. The major drawback of gene-editing technologies is off-target, which can cause unwanted editing of other genes, which hinders their wide applicability for crop trait improvement. Despite these advances, transformation and regeneration remain the major bottlenecks limiting widespread genome editing in Cannabis. Another disadvantage is the lack of efficient tissue culture methods for regeneration, transformation and regeneration of gene edited hemp crops. There are studies highlighting successful Cannabis sativa organogenesis but the commercial scale production is still a problem. However, in vitro regeneration of different varieties of hemp is very slow and found recalcitrant. This is another major disadvantage for the application of genome editing, CRISPR/Cas9 in the crop improvement of Industrial Cannabis sativa, hemp. Further limitations are caused by the regulatory uncertainty surrounding genetically edited cannabis.
Ravindra B. Malabadi, Raju K. Chalannavar· World Journal of Advanced En...· 0 citations
The application of genome editing, CRISPR/Cas9 has revolutionized plant breeding by enabling precise, efficient, and targeted modification of native genes, significantly accelerating the development of improved agronomic traits of crops. Therefore, CRISPR/Cas9 technology currently the most extensively used genome editing technique worldwide because of its simple design, cost-effectiveness, high efficiency, good reproducibility, high engineering feasibility, ability to create gene knockout, RNA editing, and quick cycle. It is used to knock in or knock out genes of interest and for generating models for genetic studies. The main components of the CRISPR/Cas9 system are an RNA-guided Cas9 endonuclease and a single-guide RNA (sgRNA). The workflow of CRISPR/Cas9 gene editing comprises selecting target sites, designing and synthesizing sgRNA, introducing transformation constructs or ribonucleoprotein (RNP) in plant cells, followed by transformation and identification of edited lines. This approach bypasses the formal regulations on GMOs, thus encouraging the widespread adoption RNA-guided gene editing in agricultural sciences and biotechnology. The system is now being utilized in the biofortification of cereal crops such as rice, wheat, barley, and maize, including vegetable crops such as potato and tomato. The world's first genome-edited rice varieties are DRR Dhan 100 (Kamala) and Pusa DST Rice 1 developed by the Indian Council of Agricultural Research (ICAR), New Delhi, India in 2025 with the objective of bringing about revolutionary changes in terms of higher production, climate adaptability, and water conservation. The CRISPR/Cas9-based crop genome editing has been utilized in imparting/producing qualitative enhancement in aroma, shelf life, sweetness, and quantitative improvement in starch, protein, gamma-aminobutyric acid (GABA), oleic acid, anthocyanin, phytic acid, gluten, and steroidal glycoalkaloid contents. Some varieties have even been modified to become disease and stress-resistant. Therefore, CRISPR/Cas9 is aiding in developing climate-ready crops and improving crop quality parameters such as appearance, palatability, nutritional components, and other preferred traits. Gene editing tools are used to generate changes to the native genetic material. Unlike GMOs, which introduce novel configurations of genetic materials typically derived from other organisms, gene editing methods modify existing genetic material in ways that can yield beneficial outcomes.
Ravindra B. Malabadi, Raju K. Chalannavar· World Journal of Advanced Re...· 0 citations
