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1.
Nat Methods ; 11(4): 393-5, 2014 Apr.
Artigo em Inglês | MEDLINE | ID: mdl-24509630

RESUMO

Firefly luciferase is the most widely used optical reporter for noninvasive bioluminescence imaging (BLI) in rodents. BLI relies on the ability of the injected luciferase substrate D-luciferin to access luciferase-expressing cells and tissues within the animal. Here we show that injection of mice with a synthetic luciferin, CycLuc1, improves BLI with existing luciferase reporters and enables imaging in the brain that could not be achieved with D-luciferin.


Assuntos
Benzotiazóis/síntese química , Medições Luminescentes/métodos , Neuroimagem/métodos , Animais , Transporte Biológico , Luciferases/metabolismo , Camundongos , Camundongos Transgênicos , Estrutura Molecular
2.
RNA ; 19(1): 96-102, 2013 Jan.
Artigo em Inglês | MEDLINE | ID: mdl-23175589

RESUMO

Alternative splicing is regulated by splicing factors that modulate splice site selection. In some cases, however, splicing factors show antagonistic activities by either activating or repressing splicing. Here, we show that these opposing outcomes are based on their binding location relative to regulated 5' splice sites. SR proteins enhance splicing only when they are recruited to the exon. However, they interfere with splicing by simply relocating them to the opposite intronic side of the splice site. hnRNP splicing factors display analogous opposing activities, but in a reversed position dependence. Activation by SR or hnRNP proteins increases splice site recognition at the earliest steps of exon definition, whereas splicing repression promotes the assembly of nonproductive complexes that arrest spliceosome assembly prior to splice site pairing. Thus, SR and hnRNP splicing factors exploit similar mechanisms to positively or negatively influence splice site selection.


Assuntos
Ribonucleoproteínas Nucleares Heterogêneas/metabolismo , Splicing de RNA/fisiologia , Éxons , Células HeLa , Ribonucleoproteínas Nucleares Heterogêneas/genética , Humanos , Íntrons , Sítios de Splice de RNA/genética , Sítios de Splice de RNA/fisiologia , Splicing de RNA/genética
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