SUMMARY RNA interference (RNAi) is a powerful research tool that has enabled molecular insights into gene activity, pathway analysis, partial loss-of-function phenotypes, and large-scale genomic discovery of gene function. While RNAi works extremely well in the non-parasitic nematode C. elegans , it is also especially useful in organisms that lack facile genetic analysis. Extensive genetic analysis of the mechanisms, delivery and regulation of RNAi in C. elegans has provided mechanistic and phenomenological insights into why RNAi is so effective in this species. These insights are useful for the testing and development of RNAi in other nematodes, including parasitic nematodes where more effective RNAi would be extremely useful. Here, we review the current advances in C. elegans for RNA delivery methods, regulation of cell autonomous and systemic RNAi phenomena, and implications of enhanced RNAi mutants. These discussions, with a focus on mechanism and cross-species application, provide new perspectives for optimizing RNAi in other species.
The administrative law literature has frequently questioned the wisdom and efficacy of generalist courts reviewing specialist agency decisionmakers. This concern has been ballooning as more and more agencies regulate activities at the frontiers of science. The courts have generally followed the substantial evidence standard enunciated in the Benzene case when reviewing scientific matters within the administrative record. In this Essay, I argue that the Benzene court inappropriately emphasizes the language of the scientific evidence when determining whether the agency has met the requisite certainty. Post-Benzene courts mandate not only a high degree of scientific certainty for decisions, but also require a high-confidence linguistic style for composing the supporting evidence. This latter requirement contradicts the writing style of the scientific community. Scientific literature uses a restrained and tentative linguistic style that the Benzene Court is unaware of, but is deeply suspicious of. Therefore, this Essay argues that close-reading of scientific literature’s language is an incorrect tool for determining scientific evidence’s certainty because it overlooks science’s linguistic tradition.
Gene knockdown by RNA interference (RNAi) in Caenorhabditis elegans is readily achieved by feeding bacteria expressing double-stranded RNA (dsRNA). Enhanced RNAi (Eri) mutants facilitate RNAi due to their hypersensitivity to dsRNA. Here, we compare eight Eri mutants for sensitivity to ingested dsRNA, targeting a variety of tissue-specific genes.
Abstract The Caenorhabditis elegans nuclear RNA interference defective (Nrde) mutants were identified by their inability to silence polycistronic transcripts in enhanced RNAi (Eri) mutant backgrounds. Here, we report additional nrde-3-dependent RNAi phenomena that extend the mechanisms, roles, and functions of nuclear RNAi. We show that nrde-3 mutants are broadly RNAi deficient and that overexpressing NRDE-3 enhances RNAi. Consistent with NRDE-3 being a dose-dependent limiting resource for effective RNAi, we find that NRDE-3 is required for eri-dependent enhanced RNAi phenotypes, although only for a subset of target genes. We then identify pgl-1 as an additional limiting RNAi resource important for eri-dependent silencing of a nonoverlapping subset of target genes, so that an nrde-3; pgl-1; eri-1 triple mutant fails to show enhanced RNAi for any tested gene. These results suggest that nrde-3 and pgl-1 define separate and independent limiting RNAi resource pathways. Limiting RNAi resources are proposed to primarily act via endogenous RNA silencing pathways. Consistent with this, we find that nrde-3 mutants misexpress genes regulated by endogenous siRNAs and incompletely silence repetitive transgene arrays. Finally, we find that nrde-3 contributes to transitive RNAi, whereby amplified silencing triggers act in trans to silence sequence-similar genes. Because nrde-dependent silencing is thought to act in cis to limit the production of primary transcripts, this result reveals an unexpected role for nuclear processes in RNAi silencing.
ABSTRACT Copyright regimes across the world share a common history and a basic modern international framework. This foundational similarity provides a unique opportunity for a comparative study of the subtler distinctions among copyright regimes. Using examples from the United States, Germany, France, Sweden, and Japan, this essay argues that American copyright law is heavily influenced by an emphasis on procedure and is much less attentive to underlying substantive rights. This essay compares and contrasts the methods by which various copyright regimes address sweat of the brow works, author moral rights, the generality of statutory provisions, and recent legislation on digital media. From these comparisons, American copyright laws seem to provide sophisticated procedural mechanisms for the copyright stakeholders as well as in copyright adjudications. For example, American courts determine whether a work is copyrightable using a largely procedural inquiry. However, such procedural means are often more burdensome than helpful in directly addressing the underlying substantive issues, like robust protection for authors' moral rights. This essay therefore highlights the theme that American copyright law places a greater emphasis on procedure than its international counterparts. More broadly, this emphasis reflects America's overarching process-centric legal values. After all, copyright laws are part of America's widely-recognized proceduralistic legal system. I. INTRODUCTION It was a cold September morning in A.D. 561. Two warring Irish clans were about to face off near modern-day Sligo County, (1) where its favorite son W.B. Yeats would be born thirteen centuries later. (2) Saint Columba was rebelling against King Diarmait for a supposedly unfair edict. (3) The ensuing Battle of Cul Dreimhne was massive and bloody, and claimed over three thousand lives. (4) This is the first known copyright dispute in history. Around this time, Christianity was spreading throughout Ireland. (5) Saint Columba, a tireless scribe who reportedly handwrote over three hundred books in his lifetime, had occasioned upon a holy psalms manuscript in a church and proceeded to transcribe a (6) The original manuscript's owner claimed ownership to Saint Columba's copy as well and appealed to King Diarmait, who announced the first copyright ruling in history: To every cow its calf and to every book its copy. (7) Saint Columba, the copyright infringer, then rebelled at the Battle of Cul Dreimhne. (8) A. Common Roots of Copyright around the World This stunning early development aside, modern day copyright law progressed through two major stages. The first copyright revolution was publisher-centric and a result of the invention of the printing press. (9) This revolution is linguistically tied to the word copyright, which implicates acts relating to copying. (10) Prior to the printing press, copying a text was laborious, expensive, and time-consuming. (11) Therefore, restricting control to the original text was a sufficient means to control the copying of the text as well. (12) With the printing press, however, mass production of information became readily available. (13) In response, many governmental and religious authorities wanted control over what was printed. (14) Thus, some of the first copyright laws were edicts that either sanctioned or prohibited the printing of specific texts. (15) For example, in 1486, the Duke of Venice sanctioned the printing of a book on the history of Venice through an exclusive grant. (16) Conversely, in 1501 Alexander VI issued a bull...against the unlicensed printing of books, and in 1559, Pope Paul IV issued a list of prohibited books that could not be printed, entitled the Index Expurgatorius. (17) Renaissance copyright was therefore a means of state censorship. (18) However, in order to balance censorship with the promotion of a publishing industry, governments began providing broader monopolistic copyright privileges as a systematic form of printing-approval. …
Small RNAs play a variety of regulatory roles, including highly conserved developmental functions. Caenorhabditis elegans not only possesses most known small RNA pathways, it is also an easy system to study their roles and interactions during development. It has been proposed that in C. elegans, some small RNA pathways compete for access to common limiting resources. The strongest evidence supporting this model is that disrupting the production or stability of endogenous short interfering RNAs (endo-siRNAs) enhances sensitivity to experimentally induced exogenous RNA interference (exo-RNAi). Here, we examine the relationship between the endo-siRNA and microRNA (miRNA) pathways, and find that, consistent with competition among these endogenous small RNA pathways, endo-siRNA pathway mutants may enhance miRNA efficacy. Furthermore, we show that exo-RNAi may also compete with both endo-siRNAs and miRNAs. Our data thus provide support that all known Dicer-dependent small RNA pathways may compete for limiting common resources. Finally, we observed that both endo-siRNA mutants and animals experiencing exo-RNAi have increased expression of miRNA-regulated stage-specific developmental genes. These observations suggest that perturbing the small RNA flux and/or the induction of exo-RNAi, even in wild-type animals, may impact development via effects on the endo-RNAi and microRNA pathways.
In Part II, I present a legal challenge to the Prescription Drug User Fee Act (PDUFA) from an administrative law perspective. While I share sympathies with those who believe PDUFA represents an unacceptable conflict of interest for the FDA, I posit arguments purely from the framework of permissible administrative agency discretion so as to avoid ambivalent analytical and empirical arguments. My argument is that given the statutory and case law determinations of permissible federal agency discretion, the FDA cannot assess a flat user fee for widely variable types of services it renders during the drug approval process. Thus, the current implementation of PDUFA is legally impermissible. Subsequently, in Part III, I compare PDUFA to three other agency user-fee mechanisms and propose specific improvements to PDFUA to minimize its conflict of interest while maintaining its revenue efficiency.
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