Silk fibres are nature’s toughest material and are of great interest for many industrial and medical applications. However, the production and spinning of artificial spider silk fibers are very challenging. Currently, the underlying mechanisms that regulate spider silk protein solubility and fiber formation remain poorly understood and artificially synthesized fibers are not as strong as the native material. We aim to generate artificial silk fibers with the same mechanical properties as native spider silk. Recently we were successful in expressing our recombinant silk protein with a 10-fold increase in yield after purification compared to previously published reports for recombinant silk proteins. In order to improve our understanding of the spinning process in the spider’s silk glands and make bioimimetic replicas thereof, it is necessary to understand which genes that are expressed in the different silk glands and to generate a spatially resolved map of the different cell types. To do this, last year we did bulk RNAseq, single cell RNAseq (ScRNAseq) for the major ampullate gland and PacBio RNA and genomic sequencing of the Swedish bridge spider, Larinioides sclopetarius. From the bulk-RNA and single cell RNA data, we succeeded in improving the annotation of the genome including the 3’UTR regions, which is required for analyzing single cell data. We also successfully sequenced the HiC data and scaffolded the genome into 13 scaffolds. These projects are ongoing, and we are analyzing the data. The outstanding quality of the data obtained so far encouraged us to initiate two new projects. The first project focusses on scRNA sequencing from four other silk glands (minor, aggregate, flagelliform and tubuliform) to further understand the molecular and cellular features within and between these silk glands. This experiment will include four biological replicates for each of the four silk glands. As a pilot project, we have successfully performed one sequencing run for each of the four silk glands (one biological replicate) libraries generated using 10x Chromium v3.1. The sequencing data from these four glands were then analyzed using cellranger and the number of cells identified in each of the gland are 7451 cells (aggregate gland), 711 cells (flagelliform gland), 1028 cells (minor gland) and 3509 cells (tubuliform gland). The next few weeks we will run single cell RNA sequencing for these four glands from three additional individuals. The second project is focused on doing in situ sequencing of the tissue sections of spider abdomen using the platform provided by 10x (former Cartana) and SciLife Lab. We have short-listed 100 genes that are identified from our single cell RNA data and that will include marker genes from different cell clusters, spidroins, carbonic anhydrase and Spider silk Complementing Elements (SpiCE) for mapping their expression in the tissue section of the spider abdomen.