Technology and Resource Development

The Zurich ORFeome Project is dedicated to the generation and distribution of well-characterized transgenic Drosophila melanogaster UAS-ORF lines. We have an expanding stock collection generated using the ΦC31 integrase method.

www.flyorf.ch

FlyORF also offers a Drosophila injection service.

www.flyorf-injection.ch

Sophisticated tools have been developed to reliably identify and trace genetically manipulated cells on the tissue level. In recent years, these technologies have been combined with transcriptomic profiling approaches down to the single cell level to reveal the molecular mechanisms that drive development, health and disease with ever increasing clarity. Yet, the substantial dropout rate is still one of the main issues of the technology: (low abundant or cell marker) transcripts, although expressed, are not detected by sequencing. A second constraint is that the sequence information retrieved for an mRNA in scRNAseq experiments cannot be preselected and is often restricted to the non-coding 3' UTR of the transcript. We have addressed these two points and have established a method for the selective "on-bead" capture of relevant cell type marker mRNAs (targeted capture). Furthermore, we can preselect and retrieve specific sequence information for selected mRNAs allowing the inspection of splice junctions, determination of recombination status of an engineered gene, detection of polymorphisms, point mutations or random mutations introduced in CRISPR/Cas9-experiments (selective sequencing). We are applying and further validating the method to specific research questions in the context of development and cancer in mouse and Drosophila and are further developing the procedures to retrieve specific sequence information on large gene panels directly within standard single cell experiments.