کد مقاله کد نشریه سال انتشار مقاله انگلیسی نسخه تمام متن
8471159 1550072 2018 34 صفحه PDF دانلود رایگان
عنوان انگلیسی مقاله ISI
The development of a sensitive fluorescent protein-based transcript reporter for high throughput screening of negative modulators of lncRNAs
ترجمه فارسی عنوان
توسعه یک خبرنگار رونویسی مبتنی بر پروتئین فلورسنت حساس برای غربالگری بالا از تعدیل کننده های منفی لنکرین
موضوعات مرتبط
علوم زیستی و بیوفناوری بیوشیمی، ژنتیک و زیست شناسی مولکولی بیولوژی سلول
چکیده انگلیسی
While the human genome is pervasively transcribed, <2% of the human genome is transcribed into protein-coding mRNAs, leaving most of the transcripts as noncoding RNAs, such as microRNAs and long-noncoding RNAs (lncRNAs), which are critical components of epigenetic regulation. lncRNAs are emerging as critical regulators of gene expression and genomic stability. However, it remains largely unknown about how lncRNAs are regulated. Here, we develop a highly sensitive and dynamic reporter that allows us to identify and/or monitor negative modulators of lncRNA transcript levels in a high throughput fashion. Specifically, we engineer a fluorescent fusion protein by fusing three copies of the PEST destruction domain of mouse ornithine decarboxylase (MODC) to the C-terminal end of the codon-optimized bilirubin-inducible fluorescent protein, designated as dBiFP, and show that the dBiFP protein is highly destabilized, compared with the commonly-used eGFP protein. We further demonstrate that the dBiFP signal is effectively down-regulated when the dBiFP and mouse lncRNA H19 chimeric transcript is silenced by mouse H19-specific siRNAs. Therefore, our results strongly suggest that the dBiFP fusion protein may serve as a sensitive and dynamic transcript reporter to monitor the inhibition of lncRNAs by microRNAs, synthetic regulatory RNA molecules, RNA binding proteins, and/or small molecule inhibitors so that novel and efficacious inhibitors targeting the epigenetic circuit can be discovered to treat human diseases such as cancer and other chronic disorders.
ناشر
Database: Elsevier - ScienceDirect (ساینس دایرکت)
Journal: Genes & Diseases - Volume 5, Issue 1, March 2018, Pages 62-74
نویسندگان
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