TY - JOUR
T1 - Methods for quantifying gene expression in ecoimmunology
T2 - From qPCR to RNA-Seq
AU - Fassbinder-Orth, Carol A.
N1 - Funding Information:
This work was supported by the Society for Integrative and Comparative Biology (DAB, DCE, DCPB) and the National Science Foundation Research Coordination Network in Ecoimmunology [NSF ISO 094177].
PY - 2014/9
Y1 - 2014/9
N2 - Synopsis Historically, the use of cutting-edge molecular techniques to study immunological gene expression and related cellular pathways has been largely limited to model organisms. Few studies have been performed that quantify the molecular immunological responses of non-model species, especially in response to environmental factors, life-history events, or exposure to parasites. This dearth of information has largely occurred due to the lack of available non-model species-specific gene sequences and immunological reagents and also due to prohibitively expensive technology. However, with the rapid development of various sequencing and transcriptomic technologies, profiling the gene expression of nonmodel organisms has become possible. Technologies and concepts explored here include an overview of current technologies for quantifying gene expression, including: qPCR, multiplex branched DNA assays, microarrays, and profiling gene expression (RNA sequencing [RNA-Seq]) based on next-generation sequencing. Examples of the advancement of these technologies in non-model systems are discussed. Additionally, applications, limitations, and feasibility of the use of these methodologies in non-model systems to address questions in ecological immunology and disease-ecology are specifically addressed.
AB - Synopsis Historically, the use of cutting-edge molecular techniques to study immunological gene expression and related cellular pathways has been largely limited to model organisms. Few studies have been performed that quantify the molecular immunological responses of non-model species, especially in response to environmental factors, life-history events, or exposure to parasites. This dearth of information has largely occurred due to the lack of available non-model species-specific gene sequences and immunological reagents and also due to prohibitively expensive technology. However, with the rapid development of various sequencing and transcriptomic technologies, profiling the gene expression of nonmodel organisms has become possible. Technologies and concepts explored here include an overview of current technologies for quantifying gene expression, including: qPCR, multiplex branched DNA assays, microarrays, and profiling gene expression (RNA sequencing [RNA-Seq]) based on next-generation sequencing. Examples of the advancement of these technologies in non-model systems are discussed. Additionally, applications, limitations, and feasibility of the use of these methodologies in non-model systems to address questions in ecological immunology and disease-ecology are specifically addressed.
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U2 - 10.1093/icb/icu023
DO - 10.1093/icb/icu023
M3 - Article
C2 - 24812328
AN - SCOPUS:84906343730
VL - 54
SP - 396
EP - 406
JO - Integrative and Comparative Biology
JF - Integrative and Comparative Biology
SN - 1540-7063
IS - 3
ER -