Advances in tools and strategies for spatiotemporal regulation of gene expression in Caenorhabditis elegans.

Spatiotemporal manipulation of gene expression is essential for dissecting gene function, elucidating developmental processes, and uncovering complex physiological mechanisms. As a model organism, Caenorhabditis elegans has played an important role in basic biology research and provides an ideal experimental system for achieving such precise control. With the development of single-copy transgenesis, CRISPR/Cas9-mediated genome editing, and conditional recombination and inducible systems, the genetic toolkits for spatiotemporal regulation of gene expression, and even at the single-cell-resolution, have been greatly expanded in C. elegans. In this review, we systematically summarize the major strategies for spatiotemporal regulation of gene expression in C. elegans, with a focus on tissue-specific RNA interference, conditional protein degradation systems, inducible transcriptional regulation systems, and recently developed approaches such as Ribo-on/Ribo-off. These methods are compared in terms of regulatory levels, technical advantages, limitations, and suitable applications. Together, these tools cover multiple layers of regulation, ranging from transcriptional and post-transcriptional control to protein-level manipulation, and provide powerful approaches for developmental biology, aging, inter-tissue signaling, and disease models, while also establishing a methodological foundation for dissecting cell-autonomous and non-cell-autonomous regulatory mechanisms in complex biological processes.