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RNA-seq analysis of MCF-10A human mammary epithelial cells with NF1 deficiency

ABSTRACT: RNA-seq analysis of MCF-10A human mammary epithelial cells with NF1 deficiency

PROVIDER: PRJNA861394 | ENA |

REPOSITORIES: ENA

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			Action	DRS
	SRR20414481_1.fastq.gz	Fastqsanger.gz
	SRR20414481_2.fastq.gz	Fastqsanger.gz
	SRR20414482_1.fastq.gz	Fastqsanger.gz
	SRR20414482_2.fastq.gz	Fastqsanger.gz
	SRR20414483_1.fastq.gz	Fastqsanger.gz

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Project description:Homologous recombination-mediated DNA repair deficiency (HRD) predisposes to cancer development, but also provides therapeutic opportunities Here, we identified an HRD gene signature that robustly predicted HRD status Unexpectedly, concurrent loss of PTEN in BRCA1-deficient cells might extensively rewire the HR repair network and confer resistance to PARP inhibitor, partially through over-expression of TTK We used the HRD gene signature as a drug discovery tool and found several PARP-inhibitor-synergizing agents through the connectivity map Thus gene expression profiling can be used to define the functional status of the HR repair network providing prognostic and therapeutic information Various shRNAs that target genes involved in homologous recombination (HR) were transfected in MCF-10A non-transformed breast cells lines Stable HR gene knockdown MCF-10A cells were seeded 200000 at 10 cm plate Cells were harvested after 48 hours culturing and used for gene expression profiling The shRNA that target Brit1 genes was transfected in MCF-10A non-transformed breast cell line by lentiviral particles and selected stable Brit1 knockdown MCF-10A cells. Scrambled control shRNA-transfected MCF-10A cells were applying as control. Both stable Brit1 knockdown and control MCF-10A cells were seeded with 2 x 10^5 cells at 10 cm culture plate. Cells were cultured in MCF-10A medium and harvested after 48 hours culturing. mRNA was extracted from collected cells and performing gene expression profiling. Three or four biological replicates were applied.

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2014-01-24 | E-GEOD-54266 | biostudies-arrayexpress

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