Fig. 4: Illustration of major applications of precise genome integration.

A In-locus tagging for gene functioning. In-locus protein tagging allows for the quantification and localization of endogenous proteins using luminescent or immunogenic tags. B Cis-regulatory element insertion. Inserting cis-regulatory elements (CREs) near promoters can modulate gene expression, leading to traits such as disease resistance and increased metabolite accumulation. C In-locus beneficial gene/allele insertion. Inserting genes or alleles at genomic safe harbor (GSH) sites enables the stable integration of beneficial traits, such as carotenoid enrichment and resistance to pathogens. These strategies are fundamental to next-generation precision breeding and synthetic biology. GT gene targeting, DOTI directional double-stranded oligonucleotide-based targeted insertion, PE prime editing, TF transcription factor, WT wild type. Created in BioRender https://BioRender.com/xqppe2r.