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Article Dans Une Revue Biochimica et Biophysica Acta:Biomembranes Année : 2006

Light controllable siRNAs regulate gene suppression and phenotypes in cells

Quan Nguyen
  • Fonction : Auteur
Rajesh Chavli
  • Fonction : Auteur
Joao Marques
  • Fonction : Auteur
  • PersonId : 2776
  • IdHAL : marquesj
Peter Conrad Ii
  • Fonction : Auteur
Die Wang
  • Fonction : Auteur
Weihai He
  • Fonction : Auteur
Barbara Belisle
  • Fonction : Auteur
Aiguo Zhang
  • Fonction : Auteur
Larry Pastor
  • Fonction : Auteur
Frank Witney
  • Fonction : Auteur
May Morris
Frederic Heitz
  • Fonction : Auteur
Gilles Divita
  • Fonction : Auteur
Bryan R.G. Williams
  • Fonction : Auteur
Gary Mcmaster
  • Fonction : Auteur

Résumé

Small interfering RNA (siRNA) is widely recognized as a powerful tool for targeted gene silencing. However, siRNA gene silencing occurs during transfection, limiting its use is in kinetic studies, deciphering toxic and off-target effects and phenotypic assays requiring temporal, and/or spatial regulation. We developed a novel controllable siRNA (csiRNA) that is activated by light. A single photo removable group is coupled during oligonucleotide synthesis to the 5' end of the antisense strand of the siRNA, which blocks the siRNA's activity. A low dose of light activates the siRNA, independent of transfection resulting in knock down of specific target mRNAs and proteins (GAPDH, p53, survivin, hNuf2) without stimulating non-specific effects such as regulated protein kinase PKR and induction of the interferon response. We demonstrate survivin and hNuf2 csiRNAs temporally knockdown their mRNAs causing multinucleation and cell death by mitotic arrest, respectively. Furthermore, we demonstrate a dose-dependent light regulation of hNuf2 csiRNA activity and resulting phenotype. The light controllable siRNAs are introduced into cells using commercially available reagents including the MPG peptide based delivery system. The csiRNAs are comparable to standard siRNAs in their transfection efficiency and potency of gene silencing. This technology should be of interest for phenotypic assays such as cell survival, cell cycle regulation, and cell development.

Dates et versions

hal-03116850 , version 1 (20-01-2021)

Identifiants

Citer

Quan Nguyen, Rajesh Chavli, Joao Marques, Peter Conrad Ii, Die Wang, et al.. Light controllable siRNAs regulate gene suppression and phenotypes in cells. Biochimica et Biophysica Acta:Biomembranes, 2006, 1758 (3), pp.394-403. ⟨10.1016/j.bbamem.2006.01.003⟩. ⟨hal-03116850⟩
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