Next Generation Sequencing Facilitates Quantitative Analysis of Wild Type, 4L;C* and Isofagamine treated 4L;C* region specifics mouse brain Transcriptomes (RNA-Seq)

Purpose: Next-generation sequencing (NGS) has revolutionized systems-based analysis of cellular pathways. The goals of this study are to compare NGS-derived brain transcriptome profiling (RNA-seq) in neuropathic region specific Gaucher mouse brain compared with WT and Isofagamine treated mice of the same age and background and secondly to identify the DEmiRNA associated with the DEmRNA before and after treatment This will give us some insights to see if miRNA is also involved in the the regulation of the expression of the genes involved in the disease process before and after treatment. Methods: 42-45 days old 4L;C*, wild-type (WT) and Isofagamine treated 4L;C* mouse brain were generated by deep sequencing, in triplicate, using IlluminaHiseq. The sequence reads that passed quality filters were analyzed at the gene level with two methods: Burrows–Wheeler Aligner (BWA) followed and TopHat followed by DESeq. qRT–PCR validation was performed using TaqMan and SYBR Green assays Overall design: Regional brain mRNA profiles of ~42 -days old wild type (WT) and 4L;C* an d Isofagamine treated mice were generated by deep sequencing, in triplicate, using IlluminaHi Seq.

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Forsberg LA, Rasi C, Pekar G, Davies H, Piotrowski A, Absher D, Razzaghian HR, Ambicka A, Halaszka K, Przewoźnik M, Kruczak A, Mandava G, Pasupulati S, Hacker J, Prakash KR, Dasari RC, Lau J, Penagos-Tafurt N, Olofsson HM, Hallberg G, Skotnicki P, Mituś J, Skokowski J, Jankowski M, Śrutek E, Zegarski W, Tiensuu Janson E, Ryś J, Tot T, Dumanski JP