ABSTRACT: There were two diversed phenotype in one T-DNA insertional line. The project is about to investigate which gene or any factor cause such diversity via genomic tools.
Project description:The membrane proteins are the main receptors that received the signals out of the cells. The abnormal of these membrane proteins will cause the wrong signal transduction which leads to loss of functions or diseases. To fix the abnormal signaling pathways, the intervention of the treatment mostly will target in these membrane receptors to inhibit the abnormal transduction cascade like the tyrosin kinase inhibitors in lung cancer. However, the lung cancer cell line have different kinds of mutation, and each of them have its own unique phenotype and drug response. The study of the membrane protein profiles from these cell lines with different mutation might help to correlate the mutation site and phenotype of the cell lines.
Project description:One of most common events in carcinogenesis is loss of DNA methylation (hypomethylation) from sequence of transposable element (TEs) including long interspersed nuclear element-1 (LINE-1). Naturally, LINE-1 and other TEs within gene body region (intragenic) will suppress by fully methylated CpG sequences in order to keep prevent incorrect transcriptional start site (TSSs) that cause genome instability. To study character of intragenic LINE-1 promoter hypomethylation or gene body methylation status, within Head and Neck squamous cell carcinoma (HNSCC) cell, LINE-1 promoter methylation status was detected with CU-L1 (unique intragenic LINE-1) and COBRALINE-1 (whole genome LINE- 1) PCRs. Each intragenic LINE-1 promoter hypomethylation have unique pattern depending on cell types and impact of LINE-1’s host gene within cell. From CU-L1 PCR 16 genes, EPHA3 and PPP2R2B are only two genes that normally induce expression by hypermethylated in gene body region, intragenic LINE-1 promoter. LINE-1 promoter hypomethylation level induce global and specific LINE-1 expression and also repress LINE-1’s host genes expression, which confirm by 5’- aza-2-deoxycytidine induced DNA hypomethylationin HNSCC cell that cause EPHA3 become greater down- regulated. Since LINE-1 RNA can express via promoter hypomethylation, knockdown LINE-1 RNA can lead increasing of EPHA3 in HNSCC cell. LINE-1 RNA was concern as key factor on LINE-1 host’s gene regulation by consequence of gene body hypomethylation. RNA molecule can regulate gene by RNAi pathway and Epigenetics mechanism, within both machinery require RISC proteins. Next, knockdown RISC protein, DICER1 gene can investigate possibility role that LINE-1 RNA regulate LINE-1 host gene. Within yeast cell, overexpress retrotransposon transcript was control by endo-siRNA pathway, which require DICER1 like protein for produce endo-siRNA from precursor molecule that include retrotransposon transcript. With CU-DREAM analyse microarray results by intersection datas for check the connection between factors on gene regulation including DICER1, LINE-1 RNA, DNA hypomethylation and HNSCC carcinogenesis model. According to CU-DREAM analysis for HNSCC cell panel, DICER1 involved pathway may induce hypermethylation on gene body region which relate to TSS and intragenic LINE-1 promoter region. While LINE-1 RNA involved pathway select to induce hypermethylation on gene without LINE-1 promoter, LINE-1 RNA have in tran function. In conclusion, intragenic LINE-1 promoter hypomethylation cause releases of LINE-1 RNA for regulate genes during HNSCC carcinogenesis via RISC protein.
Project description:Multicopy transgene arrays have enabled crucial advances in C. elegans research. Many have been in use for decades, as they are convenient tools for assaying gene expression and as cell fate reporters. Here, we show a synthetic phenotype of a multicopy molt reporter array and a missense allele, myrf-1(mg412), which was previously reported to cause a heterochronic phenotype, an additional molt in adulthood. This synthetic phenotype is due to the size or composition of the transgene. We demonstrate that additional multicopy arrays also cause synthetic phenotypes with myrf-1(mg412). Finally, using Nanopore long read sequencing, we show that additional integrated arrays all exceed the size of 6 Mb, comprising a large fraction of the nematode genome. We recommend replacing multicopy transgene reporters, whenever possible, with single- or low-copy alternatives with known insertion loci.
Project description:Multicopy transgene arrays have enabled crucial advances in C. elegans research. Many have been in use for decades, as they are convenient tools for assaying gene expression and as cell fate reporters. Here, we show a synthetic phenotype of a multicopy molt reporter array and a missense allele, myrf-1(mg412), which was previously reported to cause a heterochronic phenotype, an additional molt in adulthood. This synthetic phenotype is due to the size or composition of the transgene. We demonstrate that additional multicopy arrays also cause synthetic phenotypes with myrf-1(mg412). Finally, using Nanopore long read sequencing, we show that additional integrated arrays all exceed the size of 6 Mb, comprising a large fraction of the nematode genome. We recommend replacing multicopy transgene reporters, whenever possible, with single- or low-copy alternatives with known insertion loci.
Project description:Pinellia (Pinellia ternata (Thunb.) Breit.), as important medicinal plant, has been used to treat various ailments for a long time. The sixteen ploid plant (2n=16*13=208) Pinellia T2Plus line was obtained from an octoploid (2n=8*13=104) T2 line by chromosome-doubling technique. Compared with T2 line, the content of various medicinal components (polysaccharide, guanosine, adenosine and ephedrine) was increased in T2Plus line. In this study, the transcriptome of T2 line and T2Plus line were characterized by RNA sequencing (RNA-seq) technology. Gene Ontology (GO) and Kyoto Encyclopedia of Genes and Genomes (KEGG) pathways enrichment analysis on differential expressed unigenes (DEGs) revealed that multiple metabolic pathway were enriched significantly, such as ‘Starch and sucrose metabolism’, ‘Purine metabolism’, ‘Photosynthesis’ and six transcription factors (MYB, WRKY, bHLH, LBD lateral organ boundaries domain (LBD), HD-ZIP homeodomain-zipper (HD-ZIP) and ERF Ethylene-responsive factor (ERF) ) play a key role in difference of transcriptome between T2 line and T2Plus line. These metabolic pathways and transcription factors may play an important role in the difference of medicinal components and epigenetic features between these two Pinellia cultivars. This conclusion provides a robust theoretical basis for the mechanism of the formation of medicinal ingredients in Pinellia cultivars.