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Sci Rep ; 9(1): 9192, 2019 06 24.
Artículo en Inglés | MEDLINE | ID: mdl-31235724

RESUMEN

Introns can increase gene expression levels using a variety of mechanisms collectively referred to as Intron Mediated Enhancement (IME). IME has been measured in cell culture and plant models by quantifying expression of intronless and intron-bearing reporter genes in vitro. We developed hardware and software to implement microfluidic chip-based gene expression quantification in vivo. We altered position, number and sequence of introns in reporter genes controlled by the hsp-90 promoter. Consistent with plant and mammalian studies, we determined a single, natural or synthetic, 5'-intron is sufficient for the full IME effect conferred by three synthetic introns, while a 3'-intron is not. We found coding sequence can affect IME; the same three synthetic introns that increase mcherry protein concentration by approximately 50%, increase mEGFP by 80%. We determined IME effect size is not greatly affected by the stronger vit-2 promoter. Our microfluidic imaging approach should facilitate screens for factors affecting IME and other intron-dependent processes.


Asunto(s)
Regiones no Traducidas 5' , Caenorhabditis elegans/genética , Mejoramiento Genético , Intrones , Animales , Animales Modificados Genéticamente/genética , Expresión Génica , Perfilación de la Expresión Génica , Regulación de la Expresión Génica , Proteínas Fluorescentes Verdes/biosíntesis , Proteínas Fluorescentes Verdes/genética , Proteínas Luminiscentes/biosíntesis , Proteínas Luminiscentes/genética , Regiones Promotoras Genéticas , Proteína Fluorescente Roja
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