Targeted comparative RNA interference analysis reveals differential requirement of genes essential for cell proliferation

Yuichi Machida, Yuefeng Chen, Yuka Machida, Ankit Malhotra, Sukumar Sarkar, Anindya Dutta

Research output: Contribution to journalArticle

14 Citations (Scopus)

Abstract

Differences in the genetic and epigenetic make up of cell lines have been very useful for dissecting the roles of specific genes in the biology of a cell. Targeted comparative RNAi (TARCOR) analysis uses high throughput RNA interference (RNAi) against a targeted gene set and rigorous quantitation of the phenotype to identify genes with a differential requirement for proliferation between cell lines of different genetic backgrounds. To demonstrate the utility of such an analysis, we examined 257 growth-regulated genes in parallel in a breast epithelial cell line, MCF10A, and a prostate cancer cell line, PC3. Depletion of an unexpectedly high number of genes (25%) differentially affected proliferation of the two cell lines. Knockdown of many genes that spare PC3 (p53-) but inhibit MCF10A (p53+) proliferation induces p53 in MCF10A cells. EBNA1BP2, involved in ribosome biogenesis, is an example of such a gene, with its depletion arresting MCF10A at G1/S in a p53-dependent manner. TARCOR is thus useful for identifying cell type-specific genes and pathways involved in proliferation and also for exploring the heterogeneity of cell lines. In particular, our data emphasize the importance of considering the genetic status, when performing siRNA screens in mammalian cells.

Original languageEnglish (US)
Pages (from-to)4837-4845
Number of pages9
JournalMolecular Biology of the Cell
Volume17
Issue number11
DOIs
StatePublished - Nov 2006
Externally publishedYes

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Essential Genes
RNA Interference
Cell Proliferation
Cell Line
Genes
Gene Knockdown Techniques
Ribosomes
Epigenomics
Small Interfering RNA
Cell Biology
Prostatic Neoplasms
Breast
Epithelial Cells
Phenotype
Growth

ASJC Scopus subject areas

  • Molecular Biology
  • Genetics
  • Cell Biology

Cite this

Targeted comparative RNA interference analysis reveals differential requirement of genes essential for cell proliferation. / Machida, Yuichi; Chen, Yuefeng; Machida, Yuka; Malhotra, Ankit; Sarkar, Sukumar; Dutta, Anindya.

In: Molecular Biology of the Cell, Vol. 17, No. 11, 11.2006, p. 4837-4845.

Research output: Contribution to journalArticle

Machida, Yuichi ; Chen, Yuefeng ; Machida, Yuka ; Malhotra, Ankit ; Sarkar, Sukumar ; Dutta, Anindya. / Targeted comparative RNA interference analysis reveals differential requirement of genes essential for cell proliferation. In: Molecular Biology of the Cell. 2006 ; Vol. 17, No. 11. pp. 4837-4845.
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