RNA sequencing from epidermis of LAMA3-deficient mice

RNA sequencing data of mouse epidermis isolated from wild type and LAMA3-deficient mice Genetic, clinical and biochemical studies concurred to establish that integrity of the dermal-epidermal junction requires laminin 332, a particular subset of epithelial laminins. Laminin 332 is composed of three subunits, α3, β3 and γ2, encoded by the Lama3, Lamb3 and Lamc2 genes, respectively. In vivo functional analysis of laminin 332 in skin has been prevented because constitutive mutations of any one of the coding genes, either inherited in human or engineered in mice, cause junctional epidermolysis bullosa and early death. Consequently, it is still unknown whether and how laminin 332 contributes to skin homeostasis. To circumvent the problem, we have generated a mouse model in which disruption of the Lama3 gene is conditional and specifically induced in epidermal keratinocytes after birth. It causes a progressive depletion of laminin 332 in the skin of the mouse, which is compatible with life. To assess how laminin 332 supervises epidermal homeostasis, RNA was prepared from keratinocytes isolated from laminin 332-depleted skin and control animals (three each) to compare their gene expression profiles using RNA sequencing. Total RNA from 3 biological replicates of wild type and LAMA3-deficient mice. Paired-end sequencing 100bp reads with HiSeq2000. Lama3flox/flox mice were generated by ARTEMIS Pharmaceuticals GmbH (Cologne, Germany) by flanking exons 50 to 53 of the Lama3 gene with LoxP sites on a C57BL/6 background. Lama3flox/flox mice were crossed with keratin (K) 14 driven Cre deleter mice expressing the Cre recombinase fused to a mutated ligand-binding domain of the human estrogen receptor (K14-CreERT, Jackson laboratories), allowing temporal control of Cre activation by tamoxifen and ablation of the floxed alleles in K14-expressing basal keratinocytes. The Cre recombinase was activated with a tamoxifen-containing chow (400 mg Tamoxifen citrate/1 kg of chow). Consequently, the Lama3 gene was inactivated in the Lama3flox/flox/K14-CreERT, followed by a progressive loss of laminin 332, but not in the Lama3wt/flox / K14-CreERT used as controls. RNA was prepared from keratinocytes isolated from laminin 332-depleted skin and control animals (three mice each). Library preparation and sequencing was performed at the facilities of the Cologne Center for Genomics (University of Cologne).