High-throughput transgenesis using synthetic DNA libraries is a powerful method for systematically exploring genetic function. Diverse synthesized libraries have been used for protein engineering, identification of protein-protein interactions, characterization of promoter libraries, developmental and evolutionary lineage tracking, and various other exploratory assays. However, the need for library transgenesis has effectively restricted these approaches to single-cell models.
splits the transgenesis process into a two-step process: creation of individuals carrying experimentally introduced sequence libraries, followed by inducible extraction and integration of individual sequences/library components from the larger library cassette into engineered genomic sites. Thus, transformation of a single individual, followed by lineage expansion and functional transgenesis, gives rise to thousands of genetically unique transgenic individuals.
sites in Caenorhabditis elegans to generate a large set of individually barcoded lineages and transcriptional reporter lines from pre-defined promoter libraries. We find that this approach increases transformation yields up to ~1000-fold over current single-step methods. While we demonstrate the utility of
using C. elegans, the process is adaptable to any system where experimentally generated genomic loci landing pads and diverse, heritable DNA elements can be generated.The copyright holder for this preprint is the author/funder, who has granted bioRxiv a license to display the preprint in perpetuity.