Gene therapy offers a promising approach to treat individuals suffering from debilitating genetic diseases. Current research has uncovered a persistent and urgent need for a wider variety of promoters to support gene therapies that are tissue- and cell type-specific. Initial studies have indicated that in principle intergenic microRNA promoters (miR-p) may be a suitable source to address this requirement; however, a large and systematic effort towards isolation and functional screening of miR-ps to assess whether they may be suitable for driving gene expression has yet to be undertaken. This is in part because of a paucity of methods for the rapid and large-scale isolation of kilobase-long genomic sequences. In this project, 96 in silico predicted miR-p regions from the human genome were cloned and characterised using new high-throughput methods, including seamless gene engineering, next-generation sequencing and single-cell imaging. We found that intergenic miR-ps can drive gene expression, including promoters that have never been annotated in human genome databases. These results provide a demonstration of a next generation gene engineering platform, which will be applicable to non-coding elements more broadly, and an initial foundation of miR-ps and their ability to drive gene expression.