Legume Genomics and Genetics 2025, Vol.16, No.3, 135-142 http://cropscipublisher.com/index.php/lgg 138 MS2-based systems to activate the FLOWERING LOCUS T (FT) gene which resulted in elevated FT expression and faster flowering (Lee et al., 2019). The research on rice and tomato demonstrates that scientists can use CRISPRa and other CRISPR-based tools to modify flowering genes for time-dependent crop development (Hodaei and Werbrouck, 2023). The research shows CRISPRa technology enables scientists to study gene function and control flowering characteristics in different plant species. 4.2 Applications in legume crops (soybean, pea, related species) CRISPR applications in legumes have mainly involved gene knockout techniques yet scientists now explore CRISPRa technology for activating flowering genes through transcriptional activation. Scientists used CRISPR/Cas9 technology to modify soybean genes GmFT2a, GmPRR37 and E1 for photoperiod sensitivity and flowering time regulation to improve regional adaptation and achieve maximum yields (Hodaei and Werbrouck, 2023). The knockout methods used in these studies indicate CRISPRa technology has the potential to activate specific genes in legumes through the same regulatory networks that control flowering genes. The research demonstrates that multiple flowering-related genes can be activated through multiplexed CRISPRa systems to manage intricate traits including flowering regulation (Lee et al., 2019; Selma et al., 2022). 4.3 Mung bean flowering regulation has potential applications for translation The demonstrated success of CRISPRa in model plants and the progress in legume genome editing provide a strong foundation for applying CRISPRa to mung bean. The CRISPRa system enables scientists to control flowering time by modifying FT, CO and SOC1 regulators which results in better crop yields and environmental resistance. CRISPRa systems show increased utility for mung bean breeding and functional genomics because they can be delivered by viruses to perform simultaneous multi-gene editing (Lee et al., 2019; Selma et al., 2022). The conserved flowering pathways of mung bean with other legumes make these discoveries directly applicable for speeding up mung bean improvement through specific gene activation methods. 5 CRISPRa for Flowering Regulation in Mung Bean 5.1 Candidate flowering genes for activation in mung bean Genomic research has discovered multiple essential genes in mung bean which control the process of flowering. The Phosphatidylethanolamine-binding protein (PEBP) gene family contains FT (FLOWERING LOCUS T) clade members which function as key regulators for flower initiation. VrFT1, a member of this family, shows significantly higher expression under short-day conditions and is predicted to promote flowering, as demonstrated by overexpression studies in transgenic Arabidopsis (Xue et al., 2024). Research on PEBP genes shows that VrMFT1 and VrTFL2 and VrTFL3 genes express based on photoperiod to control the timing of flowering transition (Xue et al., 2024). The researchers have determined which genes should receive CRISPRa activation treatment to achieve optimal control of mung bean flowering duration. 5.2 Expected outcomes: early flowering, delayed flowering, adaptability CRISPRa technology allows researchers to activate VrFT1 flowering-promoting genes through CRISPRa which leads to early flowering that benefits plant growth under short cultivation periods and harsh environmental conditions. The activation of TFL-like genes which act as flowering repressors would result in delayed flowering and longer vegetative growth periods for increased yields under particular environmental conditions. The modification of these genes through fine-tuning would lead to better mung bean performance in different agro-ecological zones and result in more stable yields and breeding programs for local cultivation (Xue et al., 2024). 5.3 Technical challenges and regulatory considerations Despite the promise of CRISPRa, several technical challenges remain. The delivery of CRISPRa components to mung bean cells faces two primary obstacles which are effective delivery techniques and stable expression of dCas9-activator fusions and precise targeting of endogenous gene promoters. The solution needs to include approaches for dealing with off-target effects and the inconsistent activation performance between different gene targets and tissue types. Different countries establish separate GMO regulations for CRISPRa because it functions
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