Responsibilities include:

• Executed RNA library preparation, including TruSeq stranded total RNA, TruSeq stranded mRNA, low input mRNA, and low input total RNA
• Performed quantification by qPCR and checked for quality using Agilent TapeStation.
• Denatured sample and loaded flow cell into cartrage in sequencing instruments (NovaSeq 6000, NextSeq 500, HiSeq 4000/3000, MiSeq500/550, iSeq 100).
• De-multiplexed and converted the base-call data to fastq file.
• Analyzed data for bulk mRNA-Seq and total RNA-Seq for Bam files, FPKM value, and differential expression comparison using Python and R.
• Built up Laboratory Information Management System (LIMS) to increase work efficiency and reduce biological errors.

Project: Functional neuronal differentiation of injury-induced muscle-derived stem cell-like cells with therapeutic implications

• Injured skeletal muscle stem cells (iMuSCs) from TA muscle were isolated using the preplating method.
• Undifferentiated iMuSCs were cultured in the Neural Stem Cell medium to form the neurosphere, confirmed by immunocytochemistry and qPCR on neuronal marker expression.
• The iMuSCs in neurogenic differentiation medium could induce mature and functional neural phenotypes, evaluated by whole-cell patch-clamp recording.
• Transplantation of iMuSCs into mdx mice (a murine model representing Duchenne Muscular Dystrophy) were harvested for histological and qPCR analysis 1 and 3 weeks after injection.
• Transplantation of iMuSCs and iMuSC extract in mdx mice improved neuromuscular junction morphology by more dense aggregates and number of segments of ACHRs.

Project: Screening and characterization of small molecular inhibitors of oncogenes or genes, such as MDM2, MDMX, β-catenin both in vitro and in mice model

• Cell lines : NB1691 (mdm2 amp, p53 wt), NB1643, LA1-55N (mdm2 amp, p53 null, MYCN amp), LA1-5S (mdm2 OE, p53 null), NB1 (MYCN amp, alk amp, p53 wt), HepG2, Huh7 Hep3B, U87MG-Luc, were cultured for drug batch.
• Cell viability assay (by MTT or cell number counting), apoptosis assy, qPCR, and western blot were examined.
• xenograft, orthotopic, and patient-derived xenograft (PDX) (4~6 wks mice, NB1691-Luc), and dose and routes: 40mg/kg/day (i.p.) were applied.
• Various tissues (lungs, brain, heart, liver, spleen, GI, and kidneys) were removed, fixed, cryosectioned, (H&E) stained, and performed immunohistochemistry.
• The extracted organs and tissues were examined by qPCR and western blot.

Project: Regulation of Dendritic Development and Synaptogenesis in Hippocampal Neurons by Dlx transcription Factors

• Primary hippocampal neurons from E16.5 of mice were cultured and plated on coverslips or glass bottom dishes.
• cDNA cloning of transcriptional factor Dlx family genes and Dlx1 truncated mutants were made.
• shRNA and siRNA of downstream genes such as PAK3 and Npn-2 were constructed.
• Expression plasmids of Dlx-related cDNA, PAK3, and Npn-2, together with pGFP-N1 were cotransfected in hippocampal neuronal culture at various days in vitro.
• Inhibitory neurons, excitatory neurons, and postsynaptic marker proteins such as PSD95, GAD67, MAP2, CaMKII were fixed and immunostained
• Neuronal axon, dendrites, spine, and postsynaptic densities were captured under the epifluorescence microscope.
• Total PSD95 clusters on dendrites were measured by integrated morphometry analysis using ImageJ, and total dendrite/axon lengths were traced and quantified by Sholl analysis.