RNAi therapeutics: a potential new class of pharmaceutical drugs

Bumcrot, David; Manoharan, Muthiah; Koteliansky, Victor; Sah, Dinah W.Y.

doi:10.1038/nchembio839

Cited by 979 publications

(714 citation statements)

References 81 publications

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“…Similarly, siRNA c, which contained the causative mutation in the seed region and the silent base change at its 5' end, was not specific. From previous studies, a 5'-end mismatch within the antisense strand could have increased the potency of siRNA; 40,41 however, we observed that siRNA c, which presented such mismatches, was not more efficient than other tested siRNAs. We then compared the efficiency and specificity of siRNA d and e, in which both nucleotide changes were in the seed region, with siRNA f, in which the silent nucleotide change was in the seed region and the causative mutation was 3' of this sequence.…”

Section: Discussionmentioning

confidence: 59%

Allele-specific Col1a1 silencing reduces mutant collagen in fibroblasts from Brtl mouse, a model for classical osteogenesis imperfecta

et al. 2013

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Gene silencing approaches have the potential to become a powerful curative tool for a variety of monogenic diseases caused by gain-of-function mutations. Classical osteogenesis imperfecta (OI), a dominantly inherited bone dysplasia, is characterized in its more severe forms by synthesis of structurally abnormal type I collagen, which exerts a negative effect on extracellular matrix. Specific suppression of the mutant (Mut) allele would convert severe OI forms to the mild type caused by a quantitative defect in normal collagen. Here, we describe the in vitro and ex vivo investigation of a small interfering RNA (siRNA) approach to allele-specific gene silencing using Mut Col1a1 from the Brtl mouse, a well-characterized model for classical human OI. A human embryonic kidney cell line, which expresses the firefly luciferase gene, combined with either wild-type or Mut Brtl Col1a1 exon 23 sequences, was used for the first screening. The siRNAs selected based on their specificity and the corresponding short hairpin RNAs (shRNAs) subcloned in a lentiviral vector were evaluated ex vivo in Brtl fibroblasts for their effect on collagen transcripts and protein. A preferential reduction of the Mut allele of up to 52% was associated with about 40% decrease of the Mut protein, with no alteration of cell proliferation. Interestingly, a downregulation of HSP47, a specific collagen chaperone known to be upregulated in some OI cases, was detected. Our data support further testing of shRNAs and their delivery by lentivirus as a strategy to specifically suppress the Mut allele in mesenchymal stem cells of OI patients for autologous transplantation.

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Section: Discussionmentioning

confidence: 59%

Allele-specific Col1a1 silencing reduces mutant collagen in fibroblasts from Brtl mouse, a model for classical osteogenesis imperfecta

et al. 2013

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“…The negative charge (nearly 40 anionic charges) and size (two turns of a nucleic acid double helix) of siRNA suggests that it is unlikely to bind or cross the cell membrane unaided [51] . Much of our knowledge regarding gene packaging to facilitate siRNA delivery has come from decades of research focused on studying DNA delivery aided by cationic non-viral vectors.…”

Section: Extracellular Barriers To Sirna Deliverymentioning

confidence: 99%

Therapeutic targeting in the silent era: advances in non-viral siRNA delivery

et al. 2010

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Access to the full text of the published version may require a subscription. Rights AbstractGene silencing using RNA-interference, first described in mammalian systems almost a decade ago, is revolutionizing therapeutic target validation efforts both in vitro and in vivo. Moreover, the potential for using short interfering RNA (siRNA) as a therapy in its own right is also progressing at a significant pace. However, the widespread use of such approaches is contingent on having appropriate delivery systems to achieve clinically appropriate, safe, and efficient delivery of siRNA. There are many physicochemical and biological barriers to such delivery, and a growing emphasis on the design and characterisation of non-viral technologies that will overcome these barriers and expedite targeted delivery. This review discusses the considerations and challenges associated with use of siRNA-based therapeutics, including stability and off-target effects. Speculation is made on the properties of an ideal delivery system and the non-viral delivery approaches used to date, both in vitro and in vivo, are classified and discussed. Moreover, the ability of cyclodextrin-based delivery vectors to fulfil many of the criteria of an ideal delivery construct is also elaborated.3 Introduction:

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“…Whether miRNA-based RNAi therapeutics will develop at similar speed to siRNA therapeutics [69,70] remain to be seen. The challenges to development of siRNA-based therapeutics such as stable delivery and specificity of action [69,[71][72][73] will be revisited in the development of miRNA-based therapeutics. If these challenges can be overcome, miRNA-based therapeutics are likely to develop rapidly.…”

Section: Mirna-based Therapeuticsmentioning

confidence: 99%