کد مقاله کد نشریه سال انتشار مقاله انگلیسی نسخه تمام متن
10825868 1064681 2013 10 صفحه PDF دانلود رایگان
عنوان انگلیسی مقاله ISI
Using zebrafish transgenesis to test human genomic sequences for specific enhancer activity
موضوعات مرتبط
علوم زیستی و بیوفناوری بیوشیمی، ژنتیک و زیست شناسی مولکولی زیست شیمی
پیش نمایش صفحه اول مقاله
Using zebrafish transgenesis to test human genomic sequences for specific enhancer activity
چکیده انگلیسی
We detail an approach for the identification of human tissue-specific transcriptional enhancers involving three steps: delineation of search space around a locus or target gene, in silico identification and size definition of putative candidate sequences, and testing through several independent genomic insertions in a transgenic zebrafish reporter assay. Candidate sequences are defined through evolutionary conservation, transcription factor binding and chromatin marks (e.g. ENCODE data) and are amplified from genomic DNA, cloned into basal promoter:fluorescent protein reporter vectors based on the Tol2 transposon system and are microinjected into fertilized zebrafish eggs. After raising injected founders to sexual maturity, fluorescent screening identifies positive founder fish whose offspring undergo a detailed expression analysis to determine tissue specificity and reproducibility of specific enhancers.
ناشر
Database: Elsevier - ScienceDirect (ساینس دایرکت)
Journal: Methods - Volume 62, Issue 3, 15 August 2013, Pages 216-225
نویسندگان
, , , ,