The last decades of research in cardiovascular prevention have been characterized by successful bench-to-bedside developments for the treatment of low-density lipoprotein (LDL) hypercholesterolemia. Recent examples include the inhibition of proprotein convertase subtilisin/kexin type 9 (PCSK9) with monoclonal antibodies, small interfering RNA and antisense RNA drugs. The cumulative effects of LDL cholesterol on atherosclerosis make early, potent, and long-term reductions in LDL cholesterol desirable—ideally without the need of regular intake or application of medication and importantly, without side effects. Current reports show durable LDL cholesterol reductions in primates following one single treatment with PCSK9 gene or base editors. Use of the CRISPR/Cas system enables precise genome editing down to single-nucleotide changes. Provided safety and documentation of a reduction in cardiovascular events, this novel technique has the potential to fundamentally change our current concepts of cardiovascular prevention. In this review, the application of the CRISPR/Cas system is explained and the current state of in vivo approaches of PCSK9 editing is presented.

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"November, 1994" / Thesis (PhD)--Macquarie University, School of English & Linguistics, 1995. / Bibliography: leaves 199-212. / Introduction -- Bilingual editing of magazines -- Bilingual editing of magazines in Hong Kong -- Survey and interviews -- Grammatical bases for textual analysis -- Textual analysis -- Discussion and conclusion. / In cross-cultural or intercultural encounters of the modern age, mass communication has become a daily feature of our technological civilisation, and mass media have facilitated effective international information flow. Bilingual editing becomes an important medium of mass communication. The effectiveness of such communication rests upon the grammatical, lexical, sociolinguistic, socio-cultural, discourse and strategic competence of participants (editors, writers, translators and readers). It rests upon their ability to creatively use and to sensitively respond to language. In this dynamic process of communication, a bilingual editor not only plays the role of translator but also acts as a mediator; as Hatim and Mason (1990:223) suggest, s/he "has not only a bilingual ability but also a bi-cultural vision". -- In view of the diversity of usage of bilingual editing in the media, this research delves into the role of translation from English to Chinese in the bilingual editing of magazines in Hong Kong. This area is of interest for four reasons: first, since the press medium engages most translation practitioners, a study in this area may help future practitioners to have a better understanding of this science and art and its practice; second, text types are highly diverse, allowing room for discussion of translation devices; third, Hong Kong is a typical meeting place of the East and West and bilingual editing serves as a tool of information flow; fourth, the rising status of Chinese in Hong Kong approaching 1997 will enhance the role of bilingual editing. -- The study focuses on translation only from English and Chinese, or vice versa. In as much as there is very little academic attention to bilingual editing and its nature, processes and techniques, or to the role of translation in bilingual editing, it is believed that this research will help facilitate cross-cultural communication between Westerners and Chinese. -- The objective of this study is to derive new insights into the translation process with the support of contemporary approaches, and to descant on different lexical, grammatical and cultural features between English and Chinese; and most important, to elicit from the above features a set of parameters which may promote consistency and precision in the discussion of translating articles of the abovementioned press medium. -- Editors and theorists agree that an understanding of the source language text is essential. To review the basis for understanding the source language correctly, a text analysis of an English text and its Chinese translation will be performed. This analysis will take a functional approach which is based on Halliday's model of analysing the functional grammar of English. The first concern is with the analysis of clause complexes. The thinking behind this concern is influenced by Bell's approach to the clause. Such a functional approach is applied to the Chinese text. This does not' mean a complete application of Halliday's functional model to the Chinese language, but the functional approach will be used as a tool to reveal the relationship between the two languages as well as to analyse the source language. -- Case studies developed from the textual analysis of different types of magazines and from discussions with the translators or bilingual editors about their views of the translating process will be presented. Samples of articles illustrating the difficulties and challenges are also cited. / Mode of access: World Wide Web. / viii, 212 [28] leaves ill

Machine learning is increasingly involved in both our production and consumption of video. One symptom of this is the appearance of automated video editing applications. As this technology spreads rapidly to consumers, the need for substantive research about its social impact grows. To this end, this project maintains a focus on video editing as a microcosm of larger shifts in cultural objects co-authored by artificial intelligence. The window in which this research occurred (2010-2015) saw machine learning move increasingly into the public eye, and with it ethical concerns. What follows is, on the most abstract level, a discussion of why these ethical concerns are particularly urgent in the realm of the moving image. Algorithmic editing consists of software instructions to automate the creation of timelines of moving images. The criteria that this software uses to query a database is variable. Algorithmic authorship already exists in other media, but I will argue that the moving image is a separate case insofar as the raw material of text and music software can develop on its own. The performance of a trained actor can still not be generated by software. Thus, my focus is on the relationship between live embodied performance, and the subsequent algorithmic editing of that footage. This is a process that can employ other software like computer vision (to analyze the content of video) and predictive analytics (to guess what kind of automated film to make for a given user). How is performance altered when it has to communicate to human and non-human alike? The ritual of the iterative frame gives literal form to something that throughout human history has been a projection: the omniscient participant observer, more commonly known as the Divine. We experience black boxed software (AI's, specifically neural networks, which are intrinsically opaque) as functionally omniscient and tacitly allow it to edit more and more of life (e.g. filtering articles, playlists and even potential spouses). As long as it remains disembodied, we will continue to project the Divine on to the black box, causing cultural anxiety. In other words, predictive analytics alienate us from the source code of our cultural texts. The iterative frame then is a space in which these forces can be inscribed on the body, and hence narrated. The algorithmic editing of content is already taken for granted. The editing of moving images, in contrast, still requires a human hand. We need to understand the social power of moving image editing before it is delegated to automation. Practice Section: This project is practice-led, meaning that the portfolio of work was produced as it was being theorized. To underscore this, the portfolio comes at the end of the document. Video editors use artificial intelligence (AI) in a number of different applications, from deciding the sequencing of timelines to using facial and language detection to find actors in archives. This changes traditional production workflows on a number of levels. How can the single decision cut a between two frames of video speak to the larger epistemological shifts brought on by predictive analytics and Big Data (upon which they rely)? When predictive analytics begin modeling the world of moving images, how will our own understanding of the world change? In the practice-based section of this thesis, I explore how these shifts will change the way in which actors might approach performance. What does a gesture mean to AI and how will the editor decontextualize it? The set of a video shoot that will employ an element of AI in editing represents a move towards ritualization of production, summarized in the term the 'iterative frame'. The portfolio contains eight works that treat the set was taken as a microcosm of larger shifts in the production of culture. There is, I argue, metaphorical significance in the changing understanding of terms like 'continuity' and 'sync' on the AI-watched set. Theory Section In the theoretical section, the approach is broadly comparative. I contextualize the current dynamic by looking at previous shifts in technology that changed the relationship between production and post-production, notably the lightweight recording technology of the 1960s. This section also draws on debates in ethnographic filmmaking about the matching of film and ritual. In this body of literature, there is a focus on how participant observation can be formalized in film. Triangulating between event, participant observer and edit grammar in ethnographic filmmaking provides a useful analogy in understanding how AI as film editor might function in relation to contemporary production. Rituals occur in a frame that is dependent on a spatially/temporally separate observer. This dynamic also exists on sets bound for post-production involving AI, The convergence of film grammar and ritual grammar occurred in the 1960s under the banner of cinéma vérité in which the relationship between participant observer/ethnographer and the subject became most transparent. In Rouch and Morin's Chronicle of a Summer (1961), reflexivity became ritualized in the form of on-screen feedback sessions. The edit became transparent-the black box of cinema disappeared. Today as artificial intelligence enters the film production process this relationship begins to reverse-feedback, while it exists, becomes less transparent. The weight of the feedback ritual gets gradually shifted from presence and production to montage and post-production. Put differently, in cinéma vérité, the participant observer was most present in the frame. As participant observation gradually becomes shared with code it becomes more difficult to give it an embodied representation and thus its presence is felt more in the edit of the film. The relationship between the ritual actor and the participant observer (the algorithm) is completely mediated by the edit, a reassertion of the black box, where once it had been transparent. The crucible for looking at the relationship between algorithmic editing, participant observation and the black box is the subject in trance. In ritual trance the individual is subsumed by collective codes. Long before the advent of automated editing trance was an epistemological problem posed to film editing. In the iterative frame, for the first time, film grammar can echo ritual grammar and indeed become continuous with it. This occurs through removing the act of cutting from the causal world, and projecting this logic of post-production onto performance. Why does this occur? Ritual and specifically ritual trance is the moment when a culture gives embodied form to what it could not otherwise articulate. The trance of predictive analytics-the AI that increasingly choreographs our relationship to information-is the ineffable that finds form in the iterative frame. In the iterative frame a gesture never exists in a single instance, but in a potential state. The performers in this frame begin to understand themselves in terms of how automated indexing processes reconfigure their performance. To the extent that gestures are complicit with this mode of databasing they can be seen as votive toward the algorithmic. The practice section focuses on the poetics of this position. Chapter One focuses on cinéma vérité as a moment in which the relationship between production and post-production shifted as a function of more agile recording technology, allowing the participant observer to enter the frame. This shift becomes a lens to look at changes that AI might bring. Chapter Two treats the work of Pierre Huyghe as a 'liminal phase' in which a new relationship between production and post-production is explored. Finally, Chapter Three looks at a film in which actors perform with awareness that footage will be processed by an algorithmic edit. / The conclusion looks at the implications this way of relating to AI-especially commercial AI-through embodied performance could foster a more critical relationship to the proliferating black-boxed modes of production.

Somatische Mutationen sind eine Hauptursache für die Entstehung von Krebs. Allerdings tragen nicht alle Mutationen gleichermaßen zur Tumorentstehung bei. Ein wichtiges Ziel der personalisierten Medizin ist es daher, die für das Wachstum und Überleben des Tumors wesentlichen (sogenannte „Treiber“-Mutationen) von den zahlreichen biologisch neutralen Mutationen (sogenannte „Passagier“-Mutationen) zu unterscheiden. In der vorliegenden Studie etablierte ich einen CRISPR-basierten, genetischen Screen mit dessen Hilfe die funktionelle Rolle von Mutationen bei Krebs untersucht werden kann. Ich konnte nachweißen, dass diese mutationsselektive Strategie geeignet ist, um neue Krebstreibermutationen in der Kolorektalkarzinom- Zelllinie RKO zu identifizieren. Dazu verwendete ich 100 unterschiedliche sgRNAs, welche jeweils eine Krebsmutationssequenz spezifisch schneiden während die Wildtyp-Sequenz nicht verändert wird. Als Kontrolle nutzte ich die Kolorektalkarzinom- Zelllinie HCT116, welche die Zielmutationen nicht trägt. Interessanterweise ergab die Datenanalyse, dass zwei sgRNAs, welche die gleiche Mutation (UTP14A: S99del) schneiden, besonders rasch und ausschließlich in RKO-Zellen verloren gingen. Im Einklang mit den Screening-Ergebnissen führte die individuelle Infektion der Zellen mit diesen sgRNAs zu einem selektiven Verlust in RKO-, nicht aber HCT-Zellen, wodurch UTP14A: S99del als mutmaßliche Treiber-Mutation in RKO-Zellen identifiziert werden konnte. Die weitere Validierung und Charakterisierung dieser mutmaßlichen Treiber-Mutation wird diskutiert. Insgesamt zeigt dieser Ansatz, dass ein solches CRISPR-basiertes System ein leistungsfähiges Werkzeug auch für umfangreichere Untersuchungen von Krebsmutationen darstellt. Parallel dazu setzte ich die CRISPR-Cas-Technologie ein, um bekannte und bisher nicht therapierbare Treiber-Mutationen, wie z.B. innerhalb der Ras-Onkogen-Familie, zu untersuchen. Bemerkenswert ist in diesem Zusammenhang, dass jeder dritte Krebspatient ein durch Mutationen aktiviertes KRAS exprimiert, welches damit das am häufigsten mutierte Onkogen in menschlichen Tumorzellen ist. Im Gegensatz zu anderen Molekülen des MAPK-Signalweges konnte KRAS bisher nicht mittels kleiner, inhibitorischer Moleküle inaktiviert werden. Unter diesen Voraussetzungen birgt ein genomischer, CRISPR-basierter Ansatz das Potenzial, eine dringend benötigte therapeutische Alternative zur KRAS-Inaktivierung zu liefern. Ich entwarf daher drei mutationsselektive sgRNAs abzielend auf die häufigsten KRAS-Mutationen. Obwohl diese Strategie geeignet war, um KRAS-mutierte Tumorzellen in 3 unterschiedlichen Krebszelllinien effizient und spezifisch zu entfernen, führte die langfristige Cas9-Expression zur Bildung von onkogenen, resistenten Klonen. Dieses Phänomen wird durch DNA-Doppelstrangbrüche und die nachfolgend einsetzende, endogene DNA-Reparaturmaschinerie begünstigt. Ich konnte zeigen, dass der Adenin-Basen-Editor im Gegensatz dazu nicht nur in der Lage ist, die KRAS-Mutation ohne Doppelstrangbruch zu inaktivieren, sondern diese auch zur Wildtyp-Sequenz reparieren kann. Mit Hilfe dieses Ansatzes erreichte ich insbesondere bei Vorliegen der G12D-Mutation, einen fast vollständigen Abbau der KRAS-korrigierten Zellen. Die Validierung in patienten-abgeleiteten KRAS-G12D-Organoiden bestätigte die effiziente Korrektur sowie die daraus resultierende erhöhte Sensitivität, wenn auch in einem geringeren Maße als in Zelllinien. Somit konnte in dieser Studie erstmals gezeigt werden, dass Basen-Editierung sowohl in Zelllinien als auch in Organoiden, welche aus Tumorzellen der Patienten stammen, erfolgreich eingesetzt werden kann. Darüber hinaus ist dieses System gut verträglich und induziert weder in Zelllinien noch in Organoiden bei Vorliegen des KRAS Wildtyps unerwünschte Nebeneffekte (sogenannte „Off-target-Effekte“). Langfristig kann die Anwendung von CRISPR-basierten- und Basen-Editierungs-technologien zum Ausschalten von KRAS-Mutationen nicht nur zu einem besseren Verständnis der RAS-Biologie führen, sondern zusammen mit neuen Verabreichungsformen und Technologien die Grundlage für eine dringend benötigte KRAS-Therapie bilden.

CRISPR/Cas9 induced DNA breaks can be precisely repaired by cellular homology-directed repair (HDR) pathways using exogenously provided template DNA (donor). However, the full potential of precision editing is hindered in many model systems by low cutting efficiencies, low HDR efficiencies and, cytotoxicity related to Cas9 and donor DNA. In this thesis, I address these challenges and present methods that we developed to increase HDR efficiencies in multiple model organisms. In Caenorhabditis elegans, we show that by reducing toxicity high editing efficiencies can be achieved with single stranded oligonucleotide (ssODN) donors. We demonstrate that melting dsDNA donors dramatically improves the knock-in efficiencies of longer (1kb) edits. In addition, we describe 5′-terminal modifications to the donor molecules that further increase the frequency of precision editing. With our methodology a single optimally injected animal can yield more than 100 Green Fluorescent Protein (GFP) positive progeny, dramatically enhancing efficiency of genome editing. Next, we demonstrate the generality of 5′ modified donors by extending our studies to human cell cultures and mice zygotes. In mammalian models, 2′OMe-RNA modifications consistently increase HDR efficiencies by several fold over unmodified donors. Furthermore, end-modified donors exhibited a striking reduction in end-joining reactions including reduced concatemer formation and reduced direct ligation into the host genome. Our study demonstrates that HDR can be improved without inhibiting competing end-joining pathways and provides a platform to identify new chemical modifications that could further increase the potency and efficacy of precision genome editing.

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In this thesis, I argue that the treatment/enhancement distinction that is central to the ethical debate concerning germline genome editing and CRISPR is too vague to be ethically and normatively guiding. The problem of vagueness is twofold, being both a semantic and epistemic issue. This vagueness creates borderline cases, cases that cannot be properly defined as either treatment or enhancement, I call this The Borderline Cases Argument. These borderline cases enable a slippery slope towards eugenic practices, radical enhancement and dangerous applications of CRISPR. The distinction therefore fails to be action guiding as it cannot distinguish treatment from enhancement as well as failing to correspond to what is genuinely morally problematic with germline genome editing and not, I call this The Argument of Missing the Point. In using the treatment/enhancement distinction we therefore risk losing control over how CRISPR is used and for what purposes.

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Although peer review, in which students evaluate each others' papers, has been shown to be beneficial in many writing classrooms, the benefits of peer review to the reviewer, or the student giving the feedback, has not been thoroughly investigated in the field of second language (L2) writing. The purpose of this study is to determine which is more beneficial to improving student writing: receiving or giving peer feedback. The study was conducted at the English Language Center (ELC) at Brigham Young University (BYU). Ninety-one students in nine writing classes at two different proficiency levels, high beginning and high intermediate, participated in the study. The treatment groups reviewed anonymous papers, but received no peer feedback over the course of the semester, while the control groups received feedback, but did not review other students' papers. Writing samples collected at the beginning and end of the semester were used to evaluate which of the two methods most helped student writers. In addition, a short survey was conducted to investigate the correlation between student attitudes and demographic information and these results. Results of a series of t-tests indicated that the treatment groups, which focused solely on reviewing peers' writing, made more significant gains in their writing over the course of the semester than the control groups. These results were also more significant at the lower than the higher proficiency level. Students? level of comfort with the writing process and desire to learn how to use feedback were found to be significant predictors of these results.