A Targeted Peptide Nucleic Acid To Down-Regulate Mouse Microsomal Triglyceride Transfer Protein Expression in Hepatocytes

Rossenberg, Sabine M.W. van; Sliedregt-Bol, Karen; Prince, Perry; Berkel, Theo J.C. Van; Boom, Jacques H. van; Marel, Gijs A. van der; Biessen, Erik A.L.

doi:10.1021/bc0340417

Cited by 14 publications

(16 citation statements)

References 31 publications

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“…This negative control siRNA was selected using a modified blast to account for short sequence length and demonstrated to exclude significant homology to any known gene targets in RefSeq and MirBase (more detailed documentation on this negative control siRNA is available on the manufacturer's website [29]). Hepatocytes from female C57Black6/J mice (Charles River, Maastricht, The Netherlands) were isolated through retrograde collagenase perfusion and cultured in collagen-coated dishes exactly as previously described [30]. Twenty four hours after isolation 10 6 cells were transfected with the siRNA (final concentration 0.3, 3 or 30 nM) using the Dharmafect Duo transfection reagent® (Dharmacon, T-2010-03) according to the manufacturer's protocol.…”

Section: Methodsmentioning

confidence: 99%

Modulation of Mouse Coagulation Gene Transcription following Acute In Vivo Delivery of Synthetic Small Interfering RNAs Targeting HNF4α and C/EBPα

et al. 2012

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Hepatocyte nuclear factor 4α (HNF4α) and CCAAT/enhancer-binding protein α (C/EBPα) are important for the transcriptional control of coagulation factors. To determine in vivo the direct role of HNF4α and C/EBPα in control of genes encoding coagulation factors, a synthetic small interfering (si)RNA approach was used that enabled strong reduction of mouse hepatic HNF4α and C/EBPα under conditions that minimized target-related secondary effects. For both HNF4α and C/EBPα, intravenous injection of specific synthetic siRNAs (siHNF4α and siC/EBPα) resulted in more than 75% reduction in their liver transcript and protein levels 2 days post-injection. For siHNF4α, this coincided with marked and significantly reduced transcript levels of the coagulation genes Hrg, Proz, Serpina5, F11, F12, F13b, Serpinf2, F5, and F9 (in order of magnitude of effect) as compared to levels in control siRNA injected animals. Significant decreases in HNF4α target gene mRNA levels were also observed at 5 days post-siRNA injection, despite a limited level of HNF4α knockdown at this time point. Compared to HNF4α, C/EBPα knockdown had a modest impact on genes encoding coagulation factors. A strong reduction in C/EBPα transcript and protein levels resulted in significantly affected transcript levels of the control genes Pck1 and Fasn and a modest downregulation for coagulation genes Fba, Fbg and F5. F5 and F11 were the sole coagulation genes that were significantly affected upon prolonged (5 day) C/EBPα knockdown. We conclude that in the mouse, HNF4α has a direct and essential regulatory role for multiple hepatic coagulation genes, while a role for C/EBPα is more restricted. In addition, this study demonstrates that synthetic siRNA provides a simple and fast means for determining liver transcription factor involvement in vivo.

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Section: Methodsmentioning

confidence: 99%

Modulation of Mouse Coagulation Gene Transcription following Acute In Vivo Delivery of Synthetic Small Interfering RNAs Targeting HNF4α and C/EBPα

et al. 2012

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“…However, [ 125 I]-(GalNAc) 2 K-PNA was rapidly cleared from the blood- stream with a plasma half-life of 0.38 ± 0.04 min [211]. In another study, (GalNAc) 2 K-PNA reduced MTP expression in mouse parenchymal liver cells by 70% [212].…”

Section: Cell-penetrating Peptides Derived From Natural Proteinsmentioning

confidence: 97%

Chemical approaches to discover the full potential of peptide nucleic acids in biomedical applications

Brodyagin

Katkevičs

Kotikam

et al. 2021

Beilstein J. Org. Chem.

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Peptide nucleic acid (PNA) is arguably one of the most successful DNA mimics, despite a most dramatic departure from the native structure of DNA. The present review summarizes 30 years of research on PNA’s chemistry, optimization of structure and function, applications as probes and diagnostics, and attempts to develop new PNA therapeutics. The discussion starts with a brief review of PNA’s binding modes and structural features, followed by the most impactful chemical modifications, PNA enabled assays and diagnostics, and discussion of the current state of development of PNA therapeutics. While many modifications have improved on PNA’s binding affinity and specificity, solubility and other biophysical properties, the original PNA is still most frequently used in diagnostic and other in vitro applications. Development of therapeutics and other in vivo applications of PNA has notably lagged behind and is still limited by insufficient bioavailability and difficulties with tissue specific delivery. Relatively high doses are required to overcome poor cellular uptake and endosomal entrapment, which increases the risk of toxicity. These limitations remain unsolved problems waiting for innovative chemistry and biology to unlock the full potential of PNA in biomedical applications.

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“…PNA coupled GalNAc constructs for ASGPr targeting have also been demonstrated. By utilising PNA (PNA-K(GalNAc) 2 ) as an antisense drug directed against the microsomal triglyceride protein, a 70% down regulation of expression in isolated parenchymal liver cells was shown [60].…”

Section: Utilising the Asgp Receptor For Liver Targeted Bioplexmentioning

confidence: 99%

Bioplex Technology: Novel Synthetic Gene Delivery Pharmaceutical Based on Peptides Anchored to Nucleic Acids

Simonson

Svahn

Törnquist

et al. 2005

CPD

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Non-viral gene delivery is an important approach in order to establish safe in vivo gene therapy in the clinic. Although viral vectors currently exhibit superior gene transfer efficacy, the safety aspect of viral gene delivery is a concern. In order to improve non-viral in vivo gene delivery we have designed a pharmaceutical platform called Bioplex (biological complex). The concept of Bioplex is to link functional entities via hybridising anchors, such as Peptide Nucleic Acids (PNA), directly to naked DNA. In order to promote delivery functional entities consisting of biologically active peptides or carbohydrates, are linked to the PNA anchor. The PNA acts as genetic glue and hybridises with DNA in a sequence specific manner. By using functional entities, which elicit receptor-mediated endocytosis, improved endosomal escape and enhance nuclear entry we wish to improve the transfer of genetic material into the cell. An important aspect is that the functional entities should also have tissue-targeting properties in vivo. Examples of functional entities investigated to date are the Simian virus 40 nuclear localisation signal to improve nuclear uptake and different carbohydrate ligands in order to achieve receptor specific uptake. The delivery system is also endowed with regulatory capability, since the release of functional entities can be controlled. The aim is to create a safe, pharmaceutically defined and stable delivery system for nucleic acids with enhanced transfection properties that can be used in the clinic.

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A Targeted Peptide Nucleic Acid To Down-Regulate Mouse Microsomal Triglyceride Transfer Protein Expression in Hepatocytes

Cited by 14 publications

References 31 publications

Modulation of Mouse Coagulation Gene Transcription following Acute In Vivo Delivery of Synthetic Small Interfering RNAs Targeting HNF4α and C/EBPα

Modulation of Mouse Coagulation Gene Transcription following Acute In Vivo Delivery of Synthetic Small Interfering RNAs Targeting HNF4α and C/EBPα

Chemical approaches to discover the full potential of peptide nucleic acids in biomedical applications

Bioplex Technology: Novel Synthetic Gene Delivery Pharmaceutical Based on Peptides Anchored to Nucleic Acids

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