AON-mediated Exon Skipping Restores Ciliation in Fibroblasts Harboring the Common Leber Congenital Amaurosis CEP290 Mutation

Leber congenital amaurosis (LCA) is a severe hereditary retinal dystrophy responsible for congenital or early-onset blindness. The most common disease-causing mutation (>10%) is located deep in intron 26 of the CEP290 gene (c.2991+1655A>G). It creates a strong splice donor site that leads to insertion of a cryptic exon encoding a premature stop codon. In the present study, we show that the use of antisense oligonucleotides (AONs) allow an efficient skipping of the mutant cryptic exon and the restoration of ciliation in fibroblasts of affected patients. These data support the feasibility of an AON-mediated exon skipping strategy to correct the aberrant splicing

The N-terminal region of centrosomal protein 290 (CEP290) restores vision in a zebrafish model of human blindness

The gene coding for centrosomal protein 290 (CEP290), a large multidomain protein, is the most frequently mutated gene underlying the non-syndromic blinding disorder Leber's congenital amaurosis (LCA). CEP290 has also been implicated in several cilia-related syndromic disorders including Meckel–Gruber syndrome, Joubert syndrome, Senor–Loken syndrome and Bardet–Biedl syndrome (BBS). In this study, we characterize the developmental and functional roles of cep290 in zebrafish. An antisense oligonucleotide [Morpholino (MO)], designed to generate an altered cep290 splice product that models the most common LCA mutation, was used for gene knockdown. We show that cep290 MO-injected embryos have reduced Kupffer's vesicle size and delays in melanosome transport, two phenotypes that are observed upon knockdown of bbs genes in zebrafish. Consistent with a role in cilia function, the cep290 MO-injected embryos exhibited a curved body axis. Patients with LCA caused by mutations in CEP290 have reduced visual perception, although they present with a fully laminated retina. Similarly, the histological examination of retinas from cep290 MO-injected zebrafish revealed no gross lamination defects, yet the embryos had a statistically significant reduction in visual function. Finally, we demonstrate that the vision impairment caused by the disruption of cep290 can be rescued by expressing only the N-terminal region of the human CEP290 protein. These data reveal that a specific region of the CEP290 protein is sufficient to restore visual function and this region may be a viable gene therapy target for LCA patients with mutations in CEP290

Finishing the euchromatic sequence of the human genome

The sequence of the human genome encodes the genetic instructions for human physiology, as well as rich information about human evolution. In 2001, the International Human Genome Sequencing Consortium reported a draft sequence of the euchromatic portion of the human genome. Since then, the international collaboration has worked to convert this draft into a genome sequence with high accuracy and nearly complete coverage. Here, we report the result of this finishing process. The current genome sequence (Build 35) contains 2.85 billion nucleotides interrupted by only 341 gaps. It covers ∼99% of the euchromatic genome and is accurate to an error rate of ∼1 event per 100,000 bases. Many of the remaining euchromatic gaps are associated with segmental duplications and will require focused work with new methods. The near-complete sequence, the first for a vertebrate, greatly improves the precision of biological analyses of the human genome including studies of gene number, birth and death. Notably, the human enome seems to encode only 20,000-25,000 protein-coding genes. The genome sequence reported here should serve as a firm foundation for biomedical research in the decades ahead

Elements of a plan-based theory of speech acts

Includes bibliographical references (p. 57-61)Supported primarily by the National Research Council of Canada, and also by the National Institute of Education under Contract No. US-NIE- C-400-76-011

On the Mathematical Properties of Linguistic Theories

This paper surveys some of these results and discusses their significance for linguistic theory. However, we will avoid entirely the issue of whether one theory is more descrip- tively adequate than another. We will consider context- free, transformational, lexical-functional, generalized phrase structure, tree adjunct, and stratificational grammars) Although this paper focuses on metatheoretic results as arbiters among theories as models of human linguistic capacities, they may have other uses as well. Complexity results could be utilized for making decisions about the implementation of parsers as components of computerbased language-understanding systems. However, as Stanley Peters has pointed out, no one should underestimate "the pleasure to be derived from ferreting out these results! 3 2. Preliminary Definitions We assume that the reader is familiar with the basic defi- nitions of regular, context-free (CF), context-sensitive (CS), recursive, and recursively enumerable (r.e.) languages, as well as with their acceptors (see Hopcroft and Ullman 1979). We will be much concerned with the problem of recognizing whether a string is contained in a given language (the recognition problem) and with that of 1 This research was sponsored in part by the National Science and Engineering Research Council of Canada under Grant A9285. It was made possible in part by a gift from the Systems Development Foundation. An earlier version of this paper appeared in the Proceedings of the 21st ,4nnual Meeting of the ,4ssociation for Computational Linguistics, Cambridge, MA, June 198

Augmented Transition Networks And Their Relation To Tree Manipulation Systems.

PhDComputer scienceUniversity of Michigan, Horace H. Rackham School of Graduate Studieshttp://deepblue.lib.umich.edu/bitstream/2027.42/188824/2/7520425.pd