Molecular Soil Biology 2025, Vol.16, No.3, 114-125 http://bioscipublisher.com/index.php/msb 122 Figure 2 Schematic representation of CRISPR/Cas-based editing tools for targeting the genes related to seed quality, fiber quality, and architecture of the cotton plant. (A) DNA or RNA that encode Cas and sgRNAs or CRISPR–Cas–sgRNA ribonucleoprotein (RNP) can be transformed into the nucleus of the meristem, leaf, and root cells of a cotton plant using an appropriate delivery method such as Agrobacterium, plant viruses, polyethylene glycol (PEG), nanoparticles, and biolistic bombardment. (B) CRISPR/Cas9 comprises Cas9 endonuclease and a sgRNA complex, and a PAM site (NGG) is present downstream of the targeted DNA sequence. DSBs (blunt-ended) produced by CRISPR/Cas9 fixed by NHEJ or HDR, resulting in gene knock-out or knock-in. (C) CRISPR/Cpf1 consists of Cpf1 endonuclease and a sgRNA to bind with targeted DNA which is present upstream of a PAM site (NTT). Similarly, the DSBs (sticky-ended) generated by CRISPR/Cpf1 fix by NHEJ or HDR to gene knock out or knock-in. (D) In base editing, a base editor like adenine deaminases or cytidine deaminases is fused with nickase Cas9 (nCas9), which can lead to a base substitution in the targeted DNA sequence for gene knock-out and repair. (E) The dead Cas9 (dCas9) fused with a transcriptional repressor to regulate the targeted cotton genes associated with desired traits. (F) Epigenome editing at the target genomic site can be executed via Cas9-chromatin modifier fusion protein. (G) The dead Cas9 (dCas9) fused with the activator protein can be employed for regulating the expression of the desired genes (Adopted from Saleem et al., 2024) Current molecular breeding and genetic engineering technologies have brought new ways to improve the quality of oil cotton. Technologies such as genomics, QTL mapping, transgenics and CRISPR/Cas9 have helped us find some key functional genes, such as GhSWEET42, which plays an important role in oil synthesis and grain development. These achievements have laid the foundation for subsequent molecular design breeding and the development of new high-oil varieties. If agricultural management technologies such as precision irrigation, optimized fertilization and foliar nutrition are also combined, it will not only save resources but also improve the sustainability of cotton cultivation. It is recommended that the government invest more support in this regard, especially in basic research and new variety breeding. Consider increasing the construction of molecular breeding platforms, encouraging multidisciplinary research, and promoting the sharing and innovation of genetic resources. At the same time, it is also necessary to support the cooperation between enterprises and research institutions, and jointly promote green technologies such as water-saving irrigation and precision fertilization to enhance the competitiveness and environmental protection capabilities of the entire cotton industry chain. In the face of the challenges brought by climate change, we must also focus on those varieties that are more drought-resistant and more adaptable. In order
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