Enhancer scanning to locate regulatory regions in genomic loci.
Enhancer scanning to locate regulatory regions in genomic loci.
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DOI：
10.1038/nprot.2015.136
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发表时间：
2016-01
期刊：
影响因子：
14.8
通讯作者：
Monteiro AN
中科院分区：
文献类型：
--
作者：
Buckley M;Gjyshi A;Mendoza-Fandiño G;Baskin R;Carvalho RS;Carvalho MA;Woods NT;Monteiro AN
The present protocol provides a rapid, streamlined and scalable strategy to systematically scan genomic regions for the presence of transcriptional regulatory regions active in a specific cell type. It creates genomic tiles spanning a region of interest that are subsequently cloned by recombination into a luciferase reporter vector containing the Simian Virus 40 promoter. Tiling clones are transfected into specific cell types to test for the presence of transcriptional regulatory regions. The protocol includes testing of different SNP (single nucleotide polymorphism) alleles to determine their effect on regulatory activity. This procedure provides a systematic framework to identify candidate functional SNPs within a locus during functional analysis of genome-wide association studies. This protocol adapts and combines previous well-established molecular biology methods to provide a streamlined strategy, based on automated primer design and recombinational cloning to rapidly go from a genomic locus to a set of candidate functional SNPs in eight weeks.
