Introduction to the Gene Expression Analysis.
In 1941, Beadle and Tatum published experiments that would explain the basis of the central dogma of molecular biology, whereby the DNA through an intermediate molecule, called RNA, results proteins that perform the functions in cells. Currently, biomedical research attempts to explain the mechanisms by which develops a particular disease, for this reason, gene expression studies have proven to be a great resource. Strictly, the term "gene expression" comprises from the gene activation until the mature protein is located in its corresponding compartment to perform its function and contribute to the expression of the phenotype of cell.The expression studies are directed to detect and quantify messenger RNA (mRNA) levels of a specific gene. The development of the RNA-based gene expression studies began with the Northern Blot by Alwine et al. in 1977. In 1969, Gall and Pardue and John et al. independently developed the in situ hybridization, but this technique was not employed to detect mRNA until 1986 by Coghlan. Today, many of the techniques for quantification of RNA are deprecated because other new techniques provide more information. Currently the most widely used techniques are qPCR, expression microarrays, and RNAseq for the transcriptome analysis. In this chapter, these techniques will be reviewed.
Department of Clinical Biochemistry, University Hospital of Salamanca, Paseo de San Vicente 58, 37007, Salamanca, Spain. i.sansegundo.val@gmail.com.|Salamanca Institute for Biomedical Research (IBSAL), Salamanca, Spain. i.sansegundo.val@gmail.com.
Ignacio San
IS
Segundo-Val
Salamanca Institute for Biomedical Research (IBSAL), Salamanca, Spain.|Department of Microbiology and Genetics, University of Salamanca, Salamanca, Spain.
Catalina S
CS
Sanz-Lozano
D000818:Animals; D015870:Gene Expression; D020869:Gene Expression Profiling / Q000379:methods*; D006801:Humans; D017403:In Situ Hybridization; D020411:Oligonucleotide Array Sequence Analysis / Q000379:methods; D060888:Real-Time Polymerase Chain Reaction / Q000379:methods; D017423:Sequence Analysis, RNA / Q000379:methods
Gene expression; Microarrays; RNA sequencing; Transcriptome; qPCR