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Histone H2A as a transfection agent in crayfish hematopoietic tissue cells
Uppsala University, Disciplinary Domain of Science and Technology, Biology, Department of Physiology and Developmental Biology, Comparative Physiology.
Uppsala University, Disciplinary Domain of Science and Technology, Biology, Department of Physiology and Developmental Biology, Comparative Physiology.
2007 (English)In: Developmental and Comparative Immunology, ISSN 0145-305X, E-ISSN 1879-0089, Vol. 31, no 4, 340-346 p.Article in journal (Refereed) Published
Abstract [en]

We report a novel and highly efficient dsRNA transfection system based on one of the nuclear proteins, namely, histone H2A. RT-PCR semi-quantitative analysis of silencing target gene shows that the transfection efficiency of histone H2A is higher than Effectene or liposome-based transfection systems. Importantly, the high efficiency of histone H2A was associated with very low toxicity to the transfected crayfish hematopoietic tissue (Hpt) cells. The non-toxicity, effectiveness and specificity of histone H2A as a transfection agent provides a cheap, simple, highly efficient and reproducible gene delivery system, particularly for the sensitive cell cultures of crustacean animals such as crayfish and shrimp.

Place, publisher, year, edition, pages
2007. Vol. 31, no 4, 340-346 p.
Keyword [en]
Crayfish, Hematopoietic tissue cells, Histone H2A, RNA interference, Transfection
National Category
Biological Sciences
Identifiers
URN: urn:nbn:se:uu:diva-97409DOI: 10.1016/j.dci.2006.07.007ISI: 000244165900003PubMedID: 16999999OAI: oai:DiVA.org:uu-97409DiVA: diva2:172350
Available from: 2008-08-18 Created: 2008-08-18 Last updated: 2011-12-04
In thesis
1. Functional Studies of Some Immune Relevant Genes in a Crustacean
Open this publication in new window or tab >>Functional Studies of Some Immune Relevant Genes in a Crustacean
2008 (English)Doctoral thesis, comprehensive summary (Other academic)
Abstract [en]

The freshwater crayfish, Pacifastacus leniusculus, mounts a strong innate immune response against microbes such as viruses and bacteria. In this thesis, a novel RNA interference (RNAi) method mediated with histone H2A was developed and applied in crayfish hematopoietic tissue cell cultures for gene functional studies. Further, the interactions between host (crayfish) and pathogens (white spot syndrome virus and Aeromonas hydrophila, respectively) were studied using RNAi technology in live animals. An antilipopolysaccharide factor isolated from viral challenged crayfish by suppression subtractive hybridization was shown to interfere with the propagation of white spot syndrome virus both in vivo and in vitro in crayfish, suggesting an important role of this factor in antiviral defense. Besides, RNAi of phenoloxidase, a critical immune effector involved in melanization, revealed that phenoloxidase activity is necessary for crayfish immune defense against a highly pathogenic bacterial infection in crayfish. In addition, RNAi was also employed to study a marker protein gene involved in hemocyte maturation in crayfish. Taken together, these studies may provide more insights into the immune responses against pathogen invasion as well as hemocyte ontogenesis in crustaceans.

Place, publisher, year, edition, pages
Uppsala: Universitetsbiblioteket, 2008. 61 p.
Series
Digital Comprehensive Summaries of Uppsala Dissertations from the Faculty of Science and Technology, ISSN 1651-6214 ; 453
Keyword
anti-LPS factor, crayfish, hemocyte, innate immunity, melanization, phenoloxidase, RNA interference, WSSV
National Category
Biological Sciences
Identifiers
urn:nbn:se:uu:diva-9194 (URN)978-91-554-7252-8 (ISBN)
Public defence
2008-09-12, Lindahlsalen, EBC, Norbyvägen 18A, Uppsala, 10:00 (English)
Opponent
Supervisors
Available from: 2008-08-18 Created: 2008-08-18 Last updated: 2009-04-05Bibliographically approved

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Söderhäll, Irene

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