Characterisation of novel long non-coding RNAs in the biology of heart failure. (360G-Wellcome-093570_Z_10_A)

Initially, it was thought that just 5% of the human genome was transcribed, around one fifth of which can be accounted for by the 20,000 or so known protein-coding genes, or the messenger RNAs (1 ). However mapping and quantification of the mammalian transcriptome has shown that the number of transcripts encoded by the human genome is at least 10 times as great as the number of identified 'genes' (2, 3) that up to 70% of the mammalian genome is transcribed (4). Much of the uncharacterised transcriptors appears to be composed of long non-coding RNAs (lncRNAs), defined as RNA transcripts greater the 200nt (5), 34 of which have so far been associated with human disease (6). To begin the genome-wide search for candidate lncRNAs associated with heart failure, the Foo lab carried out RNA-seq on RNA isolated from four human control hearts and four diseased hearts. Diseased he are from male subjects, aged 40-60 years with a well-documented medical history. RNA-seq is a high throughput sequencing technique in order to obtain information about the biological system's total RNA repertoire (7). The resulting information is a summary statistic- a form of 'read count' that is linearly related to the abundance of the transcript in the sample. Figure 1 shows a diagrammatical outline of a typical RNA-seq experiment. Verify differential expression of transcripts in a larger sample of human heart tissue. Obtain complete sequence of the non-coding transcript. For the RNA-seq derived RNAs, does the transcript reside in myocytes or fibroblasts of cardiac tissue? Ensure that the transcript of interest does not yield a product of translation. What happens when the transcript is overexpressed and repressed in wild type cardiomyocytes or fibroblasts in culture? Is the Inc RNA of interest upregulated in response to stress triggers? What are the interactors of a novel Inc RNA?