RNA Interference by Single- and Double-stranded siRNA With a DNA Extension Containing a 3′ Nuclease-resistant Mini-hairpin Structure

Selective gene silencing by RNA interference (RNAi) involves double-stranded small interfering RNA (ds siRNA) composed of single-stranded (ss) guide and passenger RNAs. siRNA is recognized and processed by Ago2 and C3PO, endonucleases of the RNA-induced silencing complex (RISC). RISC cleaves passenger RNA, exposing the guide RNA for base-pairing with its homologous mRNA target. Remarkably, the 3' end of passenger RNA can accommodate a DNA extension of 19-nucleotides without loss of RNAi function. This construct is termed passenger-3'-DNA/ds siRNA and includes a 3'-nuclease-resistant mini-hairpin structure. To test this novel modification further, we have now compared the following constructs: (I) guide-3'-DNA/ds siRNA, (II) passenger-3'-DNA/ds siRNA, (III) guide-3'-DNA/ss siRNA, and (IV) passenger-3'-DNA/ss siRNA. The RNAi target was SIRT1, a cancer-specific survival factor. Constructs I-III each induced selective knock-down of SIRT1 mRNA and protein in both noncancer and cancer cells, accompanied by apoptotic cell death in the cancer cells. Construct IV, which lacks the SIRT1 guide strand, had no effect. Importantly, the 3'-DNA mini-hairpin conferred nuclease resistance to constructs I and II. Resistance required the double-stranded RNA structure since single-stranded guide-3'-DNA/ss siRNA (construct III) was susceptible to serum nucleases with associated loss of RNAi activity. The potential applications of 3'-DNA/siRNA constructs are discussed. Molecular Therapy-Nucleic Acids (2014) 2, e141; doi:10.1038/mtna.2013.68; published online 7 January 2014

Editorial

No abstract available

All things being equal? Equality and diversity in careers education,information, advice and guidance

In its education chapter, the Commission’s first Triennial Review of evidence on inequality, How Fair is Britain? Equality, Human Rights and Good Relations in 2010, found that educational attainment has been transformed in recent years. Around half of young people are now getting good qualifications at 16 (5+ A*-C GCSEs or equivalent including English and Maths) and, in 2008/09, 2.4 million students enrolled in higher education in the UK – a considerable change from a time when educational opportunities were only available to a minority of young people. However, the evidence shows that educational attainment continues to be strongly associated with socio-economic background. Stereotypical information and guidance can limit young people’s options and aspirations at an early age. Careers advice often reinforces traditional choices and young people have limited information on the pay advantages of nontraditional routes. Nearly one in four young people say that they have not had enough information to make choices for their future. This rises to just under a quarter of disabled young people

The Use of Interpreters for the Deaf and the Legal Community\u27s Obligation to Comply with the A.D.A.

Title II of the ADA, which most closely resembles section 504 of the Rehabilitation Act of 1973, requires that state and local government facilities, including courts, be accessible to individuals with disabilities. Title III of the Act requires that public accommodations be accessible to persons with disabilities. The Act specifically includes attorney\u27s offices in its definition of public accommodation. Title II and III of the Act require that reasonable accommodations be provided to qualified persons with disabilities, unless such provision would fundamentally alter the goods, services or programs provided. Reasonable accommodations can take the form of auxiliary aids and services, modifications of policies, practices and procedures, and removal of architectural barriers, to name a few. This article will focus on auxiliary aids and services appropriate to accommodating deaf and hard of hearing persons

Sovereignty is an illusion: the UK should use its power-sharing experience to play a constructive role in the EU

The UK’s ‘Westminster model’ of sovereign governance has had its day, claim Jo Murkens and Simon Toubeau. They argue that the recent transformation of the British party system and devolution have eroded the UK’s parliamentary sovereignty. The British government should learn from these developments when dealing with the EU, where sovereignty and power are also shared between the political union and its constituent parts. This is especially relevant to the Brexit discussions, since beyond the immediate horizon, the UK shares the long-term challenges of its European partners, conclude the authors

The Scottish and UK governments should beware the Ides of March

Theresa May and Nicola Sturgeon will meet today to discuss the triggering of Article 50, which will begin the process of the UK leaving the European Union. Simon Toubeau and Jo Murkens assess the likely issues up for discussion, noting that if the two leaders maintain the direction they have taken, there could be a bumpy road ahead for both Scotland and the rest of the UK

The illusion of sovereignty: the UK should embrace its experience of power-sharing at home to play a constructive role in the EU

One of the key issues in relation to the UK’s referendum is the extent to which EU membership undermines UK sovereignty. Simon Toubeau and Jo Murkens argue that this focus on sovereignty can nevertheless lead to misleading conclusions. They state that while the notion of parliamentary sovereignty lies at the heart of the Westminster model of government, this sovereignty has largely become illusionary with the growth of power-sharing and divided government within the UK. They suggest that rather than focusing on sovereignty, a better approach would be to draw on the UK’s experience of power-sharing to pursue a better relationship with the EU

Oncogenic viral protein HPV E7 up-regulates the SIRT1 longevity protein in human cervical cancer cells

Senescence is blocked in human cervical keratinocytes infected with high risk human papillomavirus (e.g. HPV type16). Viral oncoproteins HPV E6 and HPV E7 access the cell cycle via cellular p53 and retinoblastoma proteins respectively. Previously we have shown that HPV E7, not HPV E6, is also responsible for cervical cancer cell survival (SiHa cells; HPV type16). We now present evidence that SIRT1, an aging-related NAD-dependent deacetylase, mediates HPV E7 survival function in SiHa cervical cancer cells. Moreover, HPV E7 up-regulates SIRT1 protein when expressed in primary human keratinocytes. Conversely, SIRT1 levels decrease following RNAi-mediated silencing of HPV E7 in SiHa cells. Silencing HPV E6 has no effect on SIRT1 but, as expected, causes marked accumulation of p53 protein accompanied by p53-mediated up-regulation of p21. However, p53 acetylation (K382Ac) was barely detectable. Since p53 is a known SIRT1 substrate we propose that elevated SIRT1 levels (induced by HPV E7) attenuate p53 pro-apoptotic capacity via its de-acetylation. Our discovery that HPV E7 up-regulates SIRT1 links a clinically important oncogenic virus with the multi-functional SIRT1 protein. This link may open the way for a more in-depth understanding of the process of HPV-induced malignant transformation and also of the inter-relationships between aging and cancer