Comparative characterization and cytotoxicity study of TAT-peptide as potential vectors for siRNA and Dicer-substrate siRNA

Haliza Katas, Maria Abdul Ghafoor Raja, Lee Choy Ee

Research output: Contribution to journalArticle

7 Citations (Scopus)

Abstract

Recently, a newly discovered Dicer-substrate siRNA (DsiRNA) demonstrates higher potency in gene silencing than siRNA but both suffer from rapid degradation, poor cellular uptake and chemical instability. Therefore, Tat-peptide was exploited to protect and facilitate their delivery into cells. In this study, Tat-peptide was complexed with siRNA or DsiRNA through simple complexation. The physicochemical properties (particle size, surface charge and morphology) of the complexes formed were then characterized. The ability of Tat-peptide to carry and protect siRNA or DsiRNA was determined by UV-Vis spectrophotometry and serum protection assay, respectively. Cytotoxicity effect of these complexes was assessed in V79 cell line. siRNA-Tat complexes had particle size ranged from 186±17.8 to 375±8.3nm with surface charge ranged from -9.3±1.0 to +13.5±1.0mV, depending on the Tat-to-siRNA concentration ratio. As for DsiRNA-Tat complexes, the particle size was smaller than the ones complexed with siRNA, ranging from 176±8.6 to 458±14.7nm. Their surface charge was in the range of +27.1±3.6 to +38.1±0.9mV. Both oligonucleotide (ON) species bound strongly to Tat-peptide, forming stable complexes with loading efficiency of more than 86%. These complexes were relatively non cytotoxic as the cell viability of ∼90% was achieved. In conclusion, Tat-peptide has a great potential as siRNA and DsiRNA vector due to the formation of stable complexes with desirable physical characteristics, low toxicity and able to carry high amount of siRNA or DsiRNA.

Original languageEnglish
Pages (from-to)1443-1450
Number of pages8
JournalDrug Development and Industrial Pharmacy
Volume40
Issue number11
DOIs
Publication statusPublished - 1 Nov 2014

Fingerprint

Cytotoxicity
Small Interfering RNA
Peptides
Substrates
Surface charge
Particle Size
Particle size
Cells
Spectrophotometry
Gene Silencing
Complexation
Oligonucleotides
Surface morphology
Toxicity
Assays
Cell Survival
Genes

Keywords

  • Cell penetrating peptides
  • DsiRNA
  • Simple complexation
  • SiRNA
  • Tat-peptide

ASJC Scopus subject areas

  • Drug Discovery
  • Pharmacology
  • Organic Chemistry

Cite this

Comparative characterization and cytotoxicity study of TAT-peptide as potential vectors for siRNA and Dicer-substrate siRNA. / Katas, Haliza; Abdul Ghafoor Raja, Maria; Ee, Lee Choy.

In: Drug Development and Industrial Pharmacy, Vol. 40, No. 11, 01.11.2014, p. 1443-1450.

Research output: Contribution to journalArticle

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