Inhibition of RNAse A family enzymes prevents degradation and loss of silencing activity of siRNAs in serum

Haupenthal, Jörg; Baehr, Christina; Kiermayer, Simone; Zeuzem, Stefan; Piiper, Albrecht

doi:10.1016/j.bcp.2005.11.015

Cited by 133 publications

(131 citation statements)

References 39 publications

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“…25 Because inhibition of RNAse A-like enzymes in serum appears to conserve the molecular structure of the siRNA, including the 2-nucleotide 3 0 -overhangs (present study and 21 ), inhibition of RNAse A-like enzymes might reduce undesired off-target effects of siRNAs resulting from the production of blunt end dsRNA by removal of 2-nucleotide 3 0 -overhangs from siRNAs.…”

Section: Discussionmentioning

confidence: 77%

“…19 Whereas siRNA appears to be quite stable within cells, 20 our recent data indicate that siRNAs are degraded by RNAse A-like enzymes in human serum, resulting in a loss of silencing activity. 21 Our study showed that serum caused a loss of the anti-neoplastic potential of a siRNA directed against Plk1, which was prevented by inhibition of RNAse A-like enzymes.…”

mentioning

confidence: 66%

“…20 Experiments in mice may underestimate the kinetics of inactivation of siRNAs in human serum because siRNAs are more rapidly degraded in human than in mouse serum. 21 Thus, the influence of serum during a potential future human therapeutic application of siRNAs can not be estimated solely from experiments in mice. The present study investigated the influence of human serum on the anti-tumor activity of siRNA targeting the mitosis-associated serine/threonine kinase Plk1.…”

Section: Discussionmentioning

confidence: 99%

“…Screening of a number of different siRNAs targeting the mRNA of Plk1 for their ability to specifically down-modulate the expression of Plk1 in Hela cells led to the identification of a particularly effective siRNA (Plk1-siRNA1), 21 which was therefore used in the subsequent experiments.…”

Section: Plk1 Is a Suitable Target For Anti-neoplastic Therapymentioning

confidence: 99%

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RNAse A‐like enzymes in serum inhibit the anti‐neoplastic activity of siRNA targeting polo‐like kinase 1

Haupenthal

Baehr

Zeuzem

et al. 2007

Intl Journal of Cancer

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Down-modulation of target molecules in tumor cells by small interfering (si) RNAs is a promising anti-cancer strategy. A major challenge of this approach is the loss of silencing activity of the siRNAs in vivo. Our study aimed at investigating the influence of the serum compartment on the anti-tumor activity of siRNA directed against Polo-like kinase 1 (Plk1), a mitosis-associated serine/threonine kinase. The data showed that siRNA-induced suppression of Plk1 expression effectively reduced the viable cell mass and increased apoptosis in several cancer cell lines. Preincubation of the siRNA in human serum led to shortening of the siRNA as well as loss of its Plk1 silencing and anti-tumor activity. This loss of activity was prevented by inhibition of RNAse A-like enzymes. These data indicate that the anti-neoplastic effect of siRNAs declines upon incubation in human serum. This loss of anti-neoplastic activity can be prevented by inhibition of their degradation by RNAse A-like enzymes. This may have important implications for the development of a human therapeutic application of siRNAs. ' 2007 Wiley-Liss, Inc.

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Section: Discussionmentioning

confidence: 77%

mentioning

confidence: 66%

Section: Discussionmentioning

confidence: 99%

Section: Plk1 Is a Suitable Target For Anti-neoplastic Therapymentioning

confidence: 99%

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RNAse A‐like enzymes in serum inhibit the anti‐neoplastic activity of siRNA targeting polo‐like kinase 1

Haupenthal

Baehr

Zeuzem

et al. 2007

Intl Journal of Cancer

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“…Screening the efficiency of the recombinant protein siRNA delivery GAPDH siRNA on HeLa cell GAPDH. (Haupenthal et al, 2006), short serum half-life (Dykxhoorn et al, 2006;Raemdonck et al, 2008), negligible cellular internalization (Aliabadi et al, 2012), etc. The CPP, an efficient carrier for diverse cargos, could deliver siRNA via covalent conjugated or noncovalent charge interaction; however, the negatively charged siRNA can abolish the cell penetrating ability of CPP (Lee et al, 2013) via charge neutralizing.…”

Section: Discussionmentioning

confidence: 99%

Six-cell penetrating peptide-based fusion proteins for siRNA delivery

Tsui

2014

Drug Delivery

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The siRNA deliveries, for the siRNA's high negative charge, short serum half life, poor cellular internalization, etc, are still the key barriers for its application in clinic. In this study, several cell penetrating peptide (CPP) and dsRNA binding domain (dsRBD)-based fusion proteins have been developed and screened as the siRNA vector. The siRNA binding ability was measured by the agarose gel retardation, the cell uptaking was characterized under fluorescence microscopy, and further more RNAi effect was evaluated on the endogenous (GAPDH, western blot) and exogenous (GFP, flow cytometry analysis) genes in HeLa cell. Finally, the cytotoxicity was assessed on HeLa cells using cell counting kit-8. The efficiency of siRNA delivery by the CPP-dsRBD fusion protein was the CPP and the dsRBD dependent. Three fusion proteins showed similar efficiency of siRNA delivery when comparing to Lipofectamine RNAi Max as the siRNA carrier. These results indicated that these CPP-dsRBD-based fusion proteins were promising candidates as siRNA carriers.

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