Abstract: Most disease-associated genetic loci map to more than one disease or trait, suggesting they act through multiple cell types and tissues giving rise to complex disease phenotypes. This pervasive pleiotropy of human diseases presents a tremendous burden on identifying mediating mechanisms and therapeutic targets. Multiple metabolic risk haplotypes are associated with risk for metabolic diseases. However, whether a haplotype actually causes a disease and the mechanisms that cause the disease are unknown. Integration of phenotypic and transcriptional profiling in primary human cells allows for functional characterization of disease-associated genetic variants. Applicants have analyzed multiple risk haplotypes and determined the function of risk haplotypes involved in causation of specific metabolic phenotypes, such as type 2 diabetes and lipodystrophy. Methods of treatments are disclosed herein.

Abstract: Embodiments disclosed herein provide a general, scalable, high-throughput, and high-resolution approach for experimental dissection of regulatory regions and driver nucleotides in the context of human biology and disease. Applicants present HiDRA, a novel high-resolution global screen for transcriptional regulatory activity in accessible chromatin regions, enabling high-efficiency, high-throughput, and high-resolution inference of regulatory activity.

Abstract: Embodiments disclosed herein provide a general, scalable, high-throughput, and high-resolution approach for experimental dissection of regulatory regions and driver nucleotides in the context of human biology and disease. Applicants present HiDRA, a novel high-resolution global screen for transcriptional regulatory activity in accessible chromatin regions, enabling high-efficiency, high-throughput, and high-resolution inference of regulatory activity.

Abstract: Provided herein is a method for controlling a rate of fatty acid oxidation in mesenchymal cells, comprising modulating the expression or activity of ADCY5. The rate of fatty acid oxidation may be decreased by reducing the expression or activity of ADCY5. The rate of fatty acid oxidation may be increased by increasing the expression or activity of ADCY5. In some embodiments, the mesenchymal cell may be a mesenchymal stem cell, an adipocyte, an osteoblast, a chondrocyte, or a myocyte. In some embodiments, controlling the rate of fatty acid oxidation in mesenchymal stem cells comprises inhibiting fatty acid oxidation. In some embodiments, inhibiting fatty acid oxidation prevents development of Type 2 Diabetes (T2D). In some embodiments, controlling the rate of fatty acid oxidation in mesenchymal stem cells may comprise increasing the rate of fatty acid oxidation. Increasing the rate of fatty acid oxidation may promote bone formation.

Abstract: Embodiments disclosed herein provide a general, scalable, high-throughput, and high-resolution approach for experimental dissection of regulatory regions and driver nucleotides in the context of human biology and disease. Applicants present HiDRA, a novel high-resolution global screen for transcriptional regulatory activity in accessible chromatin regions, enabling high-efficiency, high-throughput, and high-resolution inference of regulatory activity.

Abstract: The invention provides homology directed repair (HDR) constructs for variant screening in cells comprising: a left and right homology arm, with either the left or right homology arm encoding a genomic edit to be incorporated at a target locus; and an excisable double selection cassette located within the left and right homology arms, the excisable double selection cassette comprising; a first selection marker; and a second selection marker; and wherein the first selection marker and the second selection marker are located between a first and second excision site. Also provided are homology directed repair (HDR) vectors comprising a construct as described herein, and methods for using such vectors.