A transposon-based chromosomal engineering method to survey a large cis-regulatory landscape in mice.

摘要: A large cis-regulatory landscape is a common feature of vertebrate genomes, particularly at key developmental gene loci with finely tuned expression patterns. Existing genetic tools for surveying genomic regions interest spanning over hundreds kilobases are limited. Here we propose chromosomal engineering strategy exploiting the local hopping trait Sleeping Beauty transposon in mouse genome. We generated embryonic stem cells targeted integration vector, carrying an enhancer-detecting lacZ reporter and loxP cassette, into developmentally critical Pax1 locus, followed by efficient transpositions, nested deletion formation derivation embryos tetraploid complementation. Comparative analysis among different insertion/deletion substantially facilitated long-range element mapping neighborhood demonstrated potential transposon-based approach as versatile tool exploration defined intervals functional or clinical relevance, such disease-associated microdeletions.