Recent advances in genome engineering technologies based on the CRISPR-associated RNA-guided endonuclease Cas9 are enabling the systematic interrogation of genome function. Analogous to the search function in modern word processors, Cas9 can be guided to specific locations within complex genomes by a short RNA search string. Using this system, DNA sequences within the endogenous genome and their functional outputs are now easily edited or modulated in virtually any organism of choice. Cas9-mediated genetic perturbation is simple and scalable, empowering researchers to elucidate the functional organization of the genome at the systems level and establish causal linkages between genetic variations and biological phenotypes. To facilitate successful and specific Cas9 targeting, we first optimize the guide RNAs (sgRNA) to significantly enhance gene editing efficiency and consistency. We also systematically characterize Cas9 targeting specificity in human cells to inform the selection of target sites and avoid off-target mutagenesis. We find that SpCas9 tolerates mismatches between guide RNA and target DNA at different positions in a sequence-dependent manner, sensitive to the number, position and distribution of mismatches. We also show that Cas9-mediated cleavage is unaffected by DNA methylation and that the dosage of Cas9 and sgRNA can be titrated to minimize off-target modification. Additionally, we provide a web-based software tool to guide the selection and validation of target sequences as well as off-target analyses. We next demonstrate that Cas9 nickase mutants can be used with paired guide RNAs to introduce targeted double-strand breaks. Because individual nicks in the genome are repaired with high fidelity, simultaneous nicking via appropriately offset guide RNAs can reduce off-target activity by over 1,500-fold in human cells. In collaboration with researchers at the University of Tokyo, we further identified a PAM-interacting domain of the Cas9 nuclease that dictates Cas9 target recognition specificity. Finally, we present protocols that provide experimentally derived guidelines for the selection of target sites, evaluation of cleavage efficiency and analysis of off-target activity. Beginning with target design, gene modifications can be achieved within as little as 1-2 weeks. Taken together, this work enables a variety of genome engineering applications from basic biology to biotechnology and medicine.

By comparing the novels of William Faulkner and Patrick White, this thesis reconsiders modernism's elitism and solipsism by revealing within them a critical interest in liberating minority perspective. Theoretical debates which continue to insist on modernism's inherent distance from the identity politics which front the postmodernist movement are overlooking modernism's deeply embedded evaluative mechanisms which work to expose and criticize the activity of psychic and social co-optation. Faulkner and White are both engaged in fictionally tracing the complexities of a failing patriarchy which can no longer substantiate its primary subjects — the white, upper class male. As representatives of modernism we can see that Faulkner and White, perhaps unwittingly, initiate the awareness that the 'failure' of their chosen subjects is in large measure due to processes of marginalization which both created the authoritative power structures within which they are constructed and helped serve to collapse them. The classic isolation of the modernist subject can be looked at not simply as an isolation predicated on endless self-referentiality, but rather on a desperate social outreaching for which he or she is not psychically equipped. By following the trajectory and perspective of specific novels and characters it becomes clear that it is precisely this handicap which clears the textual space for diversity of representation, just as it overturns the notion of modernism's functioning separatism. Chapter one concentrates on the double-edged representation of the female subject constructed as always-already 'guilty' within the psychologically, emotionally and physically repressive terms of the dominant male power structures within the context of Faulkner's Requiem for a Nun and White's A Fringe of Leaves. Chapter two investigates the psychological parameters of the morally disenfranchised modern subject whose disillusionment results from prejudicial social practices promoted by virulent racial anxiety as exemplified in Faulkner's Absalom, Absalom! and White's Voss. The third and final chapter discusses Light in August and Riders in the Chariot with attention to modernism's own investigation of the exclusion of minority voices from collective social imagining. The thesis posits that literary modernism is interested less with reconciling its literary subjects within a self-contained totalizing project than it is with invoking new social and psychological paradigms that stress the necessity of external, not internal, represented multiplicity, and that what has been (mis)recognized as modernism's self-closure is, in fact, the key not only to its own continuing relevance, but to the contemporaneous literary injunction to let all voices be heard.

Ce mémoire propose des lectures croisées de la mémoire urbaine dans Dora Bruder de Patrick Modiano et La Québécoite de Régine Robin. Les deux récits mettent en scène des narrateurs héritiers de la mémoire de la Shoah qui déambulent dans les villes de Paris et Montréal. La ville est espace d’intelligibilité dont les signes sont porteurs de sens à activer par l’observateur. À l’aide de la sémiotique de la ville (Benjamin) et des pratiques de la ville (De Certeau) et en tenant compte de la position particulière des narrateurs autour des enjeux du témoignage et de l’écriture, ce mémoire cherche à étudier comment la ville participe au déploiement d’une mémoire juive en même temps qu’elle contribue à son inévitable perte. La Deuxième Guerre mondiale a eu lieu en partie à Paris, qui en porte les traces dans une forte densité mémorielle, tandis que Montréal, ville diasporique où les événements ne se sont pas déroulés, accueille les mémoires écorchées qui se fixent d’une autre manière dans l’espace urbain. Dans les deux récits, l’espace urbain est nécessaire à la mise en texte de la rupture et de la perte, qui se dévoilent à la fois au niveau thématique (destruction urbaine, échecs répétés, perte identitaire) et formel (remise en question du récit, hybridité générique.) / This thesis offers crossed readings of urban memory in Dora Bruder from Patrick Modiano and La Québécoite from Régine Robin. Both stories depict narrators heirs of the Holocaust memory who roam the cities of Paris and Montreal. The city is a space of intelligibility whose signs are meaningful to the observer. Using the semiotics of the city (Benjamin), the practices of the city (De Certeau) and taking into account the specific position of both narrators on the issues of testimony and writing, this study seeks to explore how the city spreads the Jewish memory while at the same time contributing to its inevitable loss. The Second World War took partly place in Paris, which bears the traces in a high density of memory, while Montreal, a city where Holocaust events did not unfold, is hosting memories otherwise within its urban space. In both stories, the city is necessary to the writing of the breakdown and loss, which reveal themselves both in the background (urban destruction, repeated failures, loss of identity) and form (question of the story, generic hybridity.)

Thesis (Ph. D.)--University of Oregon, 2007. / Typescript. Includes vita and abstract. Includes bibliographical references (leaves 181-186). Also available for download via the World Wide Web; free to University of Oregon users.

Thesis (Ph. D.)--Göteborg University, 1983. / Added t.p. inserted. Includes bibliographical references (p. 194-203).

Thesis (Ph. D.)--University of Hong Kong, 2000. / Includes bibliographical references.

Zugl.: Mannheim, Univ., Diss., 2008

Zugl.: Gießen, Universiẗat, Diss. / Zugl.: Gießen, Univ., Diss.

Texte remanié de: Proefschrift--Letteren--Rijksuniversiteit te Leiden, 1996. / Bibliogr. p. 215-227. Index.

Ce mémoire vise à l´étude du processus de créolisation à partir de deux oeuvres représentatives de la littérature antillaise contemporaine: les romans Texaco, de l ´écrivain martiniquais Patrick Chamoiseau (1992) et La belle créole, de l´écrivain guadéloupéenne Maryse Condé (2001). On entreprend une étude où, à partir des concepts de créolisation (approche théorique) et figure (approche méthodologique), on examine des stratégies discursives par lesquelles le processus de créolisation a lieu dans les deux romans. Du point de vue théorique, on recourt au concept de créolisation, dégagé de textes tels que Éloge de la créolité (1989) et Écrire en pays dominé (1997), de Patrick Chamoiseau, ainsi que d´Introduction à une poétique du divers (1995), d´Édouard Glissant. Du point de vue méthodologique, ce mémoire relève des définitions de figure et imaginaire collectif de l´essaysite québécois Gérard Bouchard, surtout à partir des essais Genèse des nations et cultures du Nouveau Monde : Essai d´histoire comparée (2001) et Raison et contradiction: le mythe au secours de la pensée (2003). À travers la lecture des deux romans, on vérifie l'efficacité de chacune des trois figures – le débrouillard; l ´étranger; l´artiste/intellectuel – au moyen de lectures centrées sur une hypothèse: le processus de créolisation, sous le signe de la tension et du choc, origine d'équilibres instables, provisoires, qui surmontent (d'une façon aussi provisoire) les contradictions propres au contexte de chaque roman. Prendre la figure comme catégorie d´analyse permet ici d´apréhender la complexité de certains aspects de l´imaginaire collectif antillais. / A presente dissertação visa ao estudo da crioulização, entendida como processo, em dois romances representativos da literatura antilhana contemporânea: Texaco (1992), do escritor martinicano Patrick Chamoiseau; e La belle créole (2001), da escritora Maryse Conde, de Guadalupe. O presente estudo examina, com base nos conceitos de crioulização (abordagem teórica) e figura (abordagem metodológica), as estratégias discursivas pelas quais se dá o processo de crioulização nos dois romances. Do ponto de vista teórico, esta dissertação recorre ao conceito de crioulização, extraído de textos fundadores tais como Éloge de la créolité (1989) e Écrire en pays dominé (1997), de Patrick Chamoiseau, bem como Introduction à une poétique du Divers (1995), de Édouard Glissant. Do ponto de vista metodológico, esta dissertação é tributária das definições de figura e imaginário coletivo, formuladas pelo ensaísta quebequense Gérard Bouchard principalmente nas obras Genèse des nations et cultures du Nouveau Monde : Essai d´histoire comparée (2001) e Raison et contradiction: le mythe au secours de la pensée (2003). A partir da leitura dos dois romances, verifica-se a eficácia de cada uma das três figuras estudadas – débrouillard; étranger; artiste/intellectuel – mediante leituras centradas em uma hipótese de trabalho: o processo de crioulização, sob o signo da tensão e do choque, enseja equilíbrios instáveis, provisórios, que superam (provisoriamente) as contradições peculiares ao contexto de cada obra literária. Tomar aqui a figura como categoria de análise permite apreender a complexidade de alguns aspectos do imaginário coletivo antilhano.