Target Gene Identification and sgRNA Design for Waterlogging Tolerance in Foxtail Millet via CRISPR-Based Transcriptional Activation

Background: CRISPR activation (CRISPRa) uses non-functional Cas9 endonuclease (dCas9) but retains the genome targeting ability through its single guide RNAs (sgRNAs). CRISPRa is widely used as a gene system exploiting in recruiting various transcriptional domains (TADs) for enhancing expression of target gene(s). Drought tolerant and resource-efficient crops like millets have potential mitigating effects climate change food security. Objective: This study aimed to use Setaria italica (foxtail millet) sequence identification subsequent generation sgRNAs conferring water logging tolerance that will benefit future expansion cultivation area. Methods Results: Leveraging existing RNA-seq data information on functional studies model plants from other cereal species, maize barley, enabled candidate ERFVII foxtail millet attempt engineer tolerance. The provides step-by-step example using publicly accessible databases bioinformatics tools NCBI Phytozome characterization ortholog italica. Soft berry was promoter annotation obtain transcription start site (TSS). Subsequently, CRISP-P 2.0 design were employed generate select few efficient minimize potentially deleterious off-target binding. Conclusion: useful how advance genomics research including revolutionizing technology italica, which can be adopted plant utilization available sequence.