An efficient method for gene knock‐down by RNA interference in human skin mast cells

Mast cells (MCs) from human skin have been notoriously resistant to gene manipulation, and a method to knock‐down gene expression in in situ differentiated MCs is highly desired. The Dharmacon Accell® transfection system proved successful on several “difficult‐to‐transfect” cells. In the present work, we therefore tested this method on skin‐derived MCs using different siRNA entities. The siRNA was readily taken up, followed by pronounced, specific reduction of gene and protein expression. Hence, we present the first efficient technique for the manipulation of gene expression in primary skin MCs ex vivo, which combines high transfection rates with retained cell viability.

[1]  T. Theoharides.  Neuroendocrinology of mast cells: Challenges and controversies , 2017, Experimental dermatology.

[2]  M. Steinhoff,et al.  Integrative concepts of rosacea pathophysiology, clinical presentation and new therapeutics , 2017, Experimental dermatology.

[3]  T. Luger,et al.  Dihydroavenanthramide D inhibits mast cell degranulation and exhibits anti‐inflammatory effects through the activation of neurokinin‐1 receptor , 2017, Experimental dermatology.

[4]  T. Zuberbier,et al.  Retinoic Acid Negatively Impacts Proliferation and MCTC Specific Attributes of Human Skin Derived Mast Cells, but Reinforces Allergic Stimulability , 2017, International journal of molecular sciences.

[5]  F. Granata,et al.  Controversial role of mast cells in skin cancers , 2017, Experimental dermatology.

[6]  T. Zuberbier,et al.  IL-4 and human skin mast cells revisited: reinforcement of a pro-allergic phenotype upon prolonged exposure , 2016, Archives of Dermatological Research.

[7]  T. Zuberbier,et al.  Phenotypic variability in human skin mast cells , 2016, Experimental dermatology.

[8]  A. Seiyama,et al.  NKp46 regulates the production of serine proteases and IL‐22 in human mast cells in urticaria pigmentosa , 2015, Experimental dermatology.

[9]  T. Zuberbier,et al.  Retinoic acid potentiates inflammatory cytokines in human mast cells: Identification of mast cells as prominent constituents of the skin retinoid network , 2015, Molecular and Cellular Endocrinology.

[10]  M. Tsai,et al.  Approaches for analyzing the roles of mast cells and their proteases in vivo. , 2015, Advances in immunology.

[11]  T. Zuberbier,et al.  Skin mast cells develop non‐synchronized changes in typical lineage characteristics upon culture , 2014, Experimental dermatology.

[12]  H. Kita,et al.  Expression of Mas-related gene X2 on mast cells is upregulated in the skin of patients with severe chronic urticaria. , 2014, The Journal of allergy and clinical immunology.

[13]  T. Zuberbier,et al.  Mast cell transcriptome elucidation: what are the implications for allergic disease in the clinic and where do we go next? , 2014, Expert review of clinical immunology.

[14]  Piero Carninci,et al.  Redefinition of the human mast cell transcriptome by deep-CAGE sequencing. , 2014, Blood.

[15]  F. Niyonsaba,et al.  Critical Roles for PU.1, GATA1, and GATA2 in the Expression of Human FcεRI on Mast Cells: PU.1 and GATA1 Transactivate FCER1A, and GATA2 Transactivates FCER1A and MS4A2 , 2014, The Journal of Immunology.

[16]  B. Gibbs,et al.  Gene silencing approaches in mast cells and primary human basophils. , 2014, Methods in molecular biology.

[17]  G. Nilsson,et al.  Anti-Apoptotic Bfl-1 Is the Major Effector in Activation-Induced Human Mast Cell Survival , 2012, PloS one.

[18]  H. Nakajima,et al.  A rapid, targeted, neuron-selective, in vivo knockdown following a single intracerebroventricular injection of a novel chemically modified siRNA in the adult rat brain. , 2012, Journal of biotechnology.

[19]  F. Niyonsaba,et al.  Involvement of PU.1 in mast cell/basophil-specific function of the human IL1RL1/ST2 promoter. , 2012, Allergology international : official journal of the Japanese Society of Allergology.

[20]  T. Zuberbier,et al.  Mast cell–derived TNF‐α and histamine modify IL‐6 and IL‐8 expression and release from cutaneous tumor cells , 2011, Experimental dermatology.

[21]  P. Hokland,et al.  Nonviral transfection of leukemic primary cells and cells lines by siRNA-a direct comparison between Nucleofection and Accell delivery. , 2011, Experimental hematology.

[22]  T. Kawakami,et al.  Mast cells in atopic dermatitis. , 2009, Current opinion in immunology.

[23]  J. Schroeder,et al.  Histamine‐releasing factor/translationally controlled tumor protein (HRF/TCTP)‐induced histamine release is enhanced with SHIP‐1 knockdown in cultured human mast cell and basophil models , 2008, Journal of leukocyte biology.

[24]  G. Nilsson,et al.  Is there a role for mast cells in psoriasis? , 2008, Archives of Dermatological Research.

[25]  D. Metcalfe,et al.  The mast cell and allergic diseases: role in pathogenesis and implications for therapy , 2007, Clinical and experimental allergy : journal of the British Society for Allergy and Clinical Immunology.

[26]  A. Kaplan,et al.  Progress and Challenges in the Understanding of Chronic Urticaria , 2007, Allergy, asthma, and clinical immunology : official journal of the Canadian Society of Allergy and Clinical Immunology.

[27]  M. Nishiyama,et al.  FOG-1 represses GATA-1-dependent FcϵRI β-chain transcription: transcriptional mechanism of mast-cell-specific gene expression in mice , 2006 .

[28]  M. Nishiyama,et al.  FOG-1 represses GATA-1-dependent FcepsilonRI beta-chain transcription: transcriptional mechanism of mast-cell-specific gene expression in mice. , 2006, Blood.

[29]  T. Zuberbier,et al.  The transcription factor profile of human mast cells in comparison with monocytes and granulocytes , 2004, Cellular and Molecular Life Sciences CMLS.

[30]  R. Stevens,et al.  RasGRP4 Regulates the Expression of Prostaglandin D2in Human and Rat Mast Cell Lines* , 2003, The Journal of Biological Chemistry.