Legume Genomics and Genetics 2025, Vol.16, No.3, 135-142 http://cropscipublisher.com/index.php/lgg 139 as a tool that produces no permanent DNA changes. The successful implementation of CRISPRa-modified mung bean varieties depends on obtaining public approval and establishing precise regulatory frameworks for their use in agriculture. 5 CRISPRa for Flowering Regulation in Mung Bean 5.1 Candidate flowering genes for activation in mung bean Research through genomic analysis has discovered multiple essential genes in mung bean which control the process of flowering. The Phosphatidylethanolamine-binding protein (PEBP) gene family contains the FT (FLOWERING LOCUS T) clade which functions as a key regulator for flower initiation. The family contains VrFT1 which shows increased expression under short-day conditions and studies with transgenic Arabidopsis overexpression demonstrate its role in promoting flowering. The study of PEBP genes progresses through VrMFT1 and VrTFL2 and VrTFL3 which show photoperiod-dependent expression to control flowering time changes (Xue et al., 2024). The researchers have determined which genes should receive CRISPRa activation to achieve the best results for mung bean flowering duration control. 5.2 Expected outcomes: early flowering, delayed flowering, adaptability The activation of VrFT1 flowering-promoting genes through CRISPRa technology will lead to early flowering which benefits plant adaptation to brief cultivation periods and harsh environmental conditions. The activation of TFL-like genes which function as flowering repressors presents a method to postpone flowering which could enhance plant growth and production in specific situations. The modification of these genes through fine-tuning would allow mung bean to grow in different agricultural areas and make breeding for local cultivation easier (Xue et al., 2024). 5.3 Technical challenges and regulatory considerations Despite the promise of CRISPRa, several technical challenges remain. CRISPRa technology encounters two major challenges which include delivering CRISPRa components into mung bean cells and sustaining dCas9-activator fusion expression stability and precise endogenous gene promoter targeting. The research requires solutions to handle off-target effects and variable activation efficiency that affects different gene targets and tissues. The classification of CRISPRa under GMO regulations varies across countries because it functions as a tool which makes no permanent changes to DNA sequences. CRISPRa-modified mung bean varieties need public approval together with specific regulatory frameworks to succeed in agricultural applications. 6 Case Study: Activating VrFT1 Homolog in Mung Bean 6.1 Experimental design: activating VrFT1 homolog in mung bean The VrFT1 gene, a member of the Phosphatidylethanolamine-binding protein (PEBP) family, has been identified as a key regulator of flowering in mung bean. Scientists studied PEBP gene function through genome-wide analysis of these genes and studied their expression patterns under various photoperiod conditions. The study of VrFT1 activation through CRISPRa in mung bean remains unreported but researchers used transgenic Arabidopsis to study VrFT1 function as a substitute for mung bean gene activation studies (Xue et al., 2024). The experimental design measured VrFT1 expression levels across different tissues and under short-day (SD) and long-day (LD) environmental conditions before researchers studied the model plant system for phenotypic effects. 6.2 Observed results: changes in flowering time and yield traits The research established that VrFT1 expression levels reached their peak under SD conditions which resulted in plants flowering at an earlier stage. Xue et al. (2024) proved VrFT1 functions as a flowering promoter through their study which showed that Arabidopsis plants with VrFT1 overexpression reached flowering stage before control plants did. The VrFT1 and PEBP genes in mung bean showed photoperiod-dependent and tissue-specific expression patterns which indicates their function in flowering time regulation. The study does not include direct measurements of mung bean yield traits but suggests that VrFT1 overexpression enables researchers to control flowering time for better yield performance under different environmental conditions (Figure 2).
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