Líffræðifélag Íslands
Líffræðiráðstefnan 2015
Erindi/veggspjald / Talk/poster V69
Sreekala Syamala Kumary (1), Ottar Rolfsson (1), Steinn Gudmundsson(1)
1. Center fyrir Systems Biology,Háskóli Íslands
Kynnir / Presenter: Sreekala Syamala Kumary
Tengiliður / Corresponding author: Ottar Rolfsson (ottarr@hi.is )
Introduction: Methods for functional characterization of missing biological components in prokaryotes is well established but their application in eukaryotes has been more limited. In this context, we are exploring a novel systems biology approach for the functional characterization of human genes and their protein products through computational metabolic gap filling analysis followed by their experimental validation. Methods and data: BLAST analysis was done to infer the functions of uncharacterized proteins and the predicted functions were added to human metabolic reconstruction network Recon 2. Metabolic network gap analysis and COBRA were done to computationally evaluate the ability of these genes to fill gaps in Recon 2. Results: We have identified 425 uncharacterized human proteins with catalytic activity from UniProt database based on their gene ontology profiles and proposed their metabolic roles using sequence homology methods. After adding them to Recon 2, 13 gene targets were selected using COBRA. Specific CRISPR gRNA expression plasmids of these genes were generated by cloning designed CRISPR guide RNAs into BsmBI digested MLM3636 vector and their knock out and metabolomic studies are progressing in human Hep g2 cells. Conclusions: We have been successful in predicting and validating the functions of uncharacterized proteins by connecting them to Recon 2. The method represents a novel approach to propose metabolic functions encoded for in the human genome.