Unknown,Transcriptomics,Genomics,Proteomics

Dataset Information

Transcription profiling of human HeLa cells with intended transcriptional silencing with siRNAs siVFp(-992) and siVFp(-992)m1.

ABSTRACT: Microarray analysis of gene expression profile human cell line (HeLa) treated with siRNA.

ORGANISM(S): Homo sapiens

SUBMITTER: Joshua Moses

PROVIDER: E-MEXP-2221 | biostudies-arrayexpress |

REPOSITORIES: biostudies-arrayexpress

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Publications

Intended transcriptional silencing with siRNA results in gene repression through sequence-specific off-targeting.

Moses Joshua J Goodchild Amber A Rivory Laurent P LP

RNA (New York, N.Y.) 20091221 2

Transcriptional gene silencing has been reported with siRNA targeting the promoter region of genes. We tested several siRNAs directed against the human VEGF promoter. Of these, siVFp(-992) exhibited > or =50% suppression of VEGF production in two human cell lines. To determine the specificity of this siRNA-mediated suppression, plasmids were prepared to express a luciferase reporter under the control of VEGF promoters featuring wild-type, mutated, or deleted target sequences. siRNA transfection ...[more]

PMID: 20026621

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Project description:Twist1, a basic helix-loop-helix transcription factor, is expressed in mesenchymal precursor populations during embryogenesis and in metastatic cancer cells. In the developing heart, Twist1 is highly expressed in endocardial cushion (ECC) valve mesenchymal cells and is down regulated during valve differentiation and remodeling. Previous studies demonstrated that Twist1 promotes cell proliferation, migration, and expression of primitive ECM molecules in ECC mesenchymal cells. Furthermore, Twist1 expression is induced in human pediatric and adult diseased heart valves. However, the Twist1 downstream target genes that mediate increased cell proliferation and migration during early heart valve development remain largely unknown. Candidate gene and global gene profiling approaches were used to identify direct transcriptional targets of Twist1 during heart valve development. Candidate target genes were analyzed for evolutionarily conserved regions (ECRs) containing E-box consensus sequences that are potential Twist1 binding sequences. ECRs containing conserved E-box sequences were identified for Twist1 responsive genes Tbx20, Cdh11, Sema3C, Rab39b, and Gadd45a. Twist1 binding to these sequences in vivo was determined by chromatin immunoprecipitation assays, and binding was detected in ECCs but not late stage remodeling valves. In addition identified Twist1 target genes are highly expressed in ECCs and have reduced expression during heart valve remodeling in vivo which is consistent with the expression pattern of Twist1. Together these analyses identify multiple new genes involved in cell proliferation and migration that are differentially expressed in the developing heart valves, are responsive to Twist1 transcriptional function, and contain Twist1 responsive regulatory sequences. Murine MC3T3-E1 preosteoblast cells were treated with siScrambled control or siTwist1 to achieve knockdown of Twist1. Both siScr and siTwist1 were performed in triplicate. Isolated RNA was analyzed with Affymetrix muring MOE 430 2.0 gene chip microarray.

Dataset Information

Transcription profiling of human HeLa cells with intended transcriptional silencing with siRNAs siVFp(-992) and siVFp(-992)m1.

Publications

Intended transcriptional silencing with siRNA results in gene repression through sequence-specific off-targeting.

Similar Datasets

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