Biophysical Society Thematic Meeting - November 16-20, 2015

Biophysics in the Understanding, Diagnosis, and Treatment of Infectious Diseases Poster Abstracts

24-POS Board 24 LncRNA Discovery in Host-Pathogen Interactions Loretta Magagula 1,2,3 , Janine Scholefield 1,2 , Youtaro Shibayama 1,2 , Joana Cruz 4 , Frank Brombacher 3 , Musa Mhlanga 2,3,4 . 1 Council of Scientific & Industrial Research (CSIR), Pretoria, Gauteng, South Africa, 2 University of Cape Town, Cape Town, Western Cape, South Africa, 4 Universidade de Lisboa, Lisboa, Portugal. 3 University of Cape Town, Cape Town, Western Cape, South Africa, In the last year alone, a handful of studies have identified long noncoding RNAs (lncRNAs) linc- Cox2, Lethe, PACER and THRIL as central molecular players in host cell innate immune response against microbial infection. Gene regulation has emerged as a prevailing theme in lncRNA functioning. These discoveries and the vast numbers of uncharacterized lncRNAs identified by high-throughput next-generation transcriptome sequencing technologies, set a precedence for further investigation and characterization of lncRNAs in infection biology. Importantly, lncRNAs may serve as important diagnostic markers of infection as well as therapeutic targets. These aspects, although extensively being explored in cancer research, have been neglected in infection biology, particularly in microbial infection. In this study, RNA-Sequencing (RNA- Seq ) was used to identify subtle vairations in transcriptional activity, with particular emphasis on lncRNA differential expression, and uncover their physiological relevance during Listeria monotocytogenes infection. To this end, an RNA- Seq dataset of Listeria -infected HeLa cells was subjected to several variations of data analysis lncRNA discovery pipelines. Potential lncRNA functioning was hypothesized using a derivation of the Rinn & Chang “guilt by association” approach in which lncRNA functioning was hypothesized based on known functions of tightly co-expressed protein coding mRNAs. “Guilty” lncRNAs were then knocked down in the HeLa cells using transcription activator-like nucleases (TALENs) to validate their candidacy as infection-regulating lncRNAs. Preliminary investigations conducted in this study have revealed potential Listeria infection-inhibiting lncRNA candidates in the HeLa cell model. Furthermore, we are currently exploring the use of a physiologically relevant cellular model, induced pluripotent stem cell (iPSC) monocyte derived macrophages (MDMs), to validate identified lncRNA candidates.

57