Publications
2022
2021
Insulin receptor substrate 1, but not IRS2, plays a dominant role in regulating pancreatic alpha cell function in mice
. J Biol Chem. 2021.A Systematic Comparison of Protocols for Recovery of High-Quality RNA from Human Islets Extracted by Laser Capture Microdissection
. Biomolecules. 2021.Differential roles of FOXO transcription factors on insulin action in brown and white adipose tissue
. J Clin Invest . 2021.Essential roles of insulin and IGF-1 receptors during embryonic lineage development
. Mol Metab . 2021.Harnessing reaction-based probes to preferentially target pancreatic β-cells and β-like cells
. Life Science Alliance . 2021.Defective insulin receptor signaling in hPSCs skews pluripotency and negatively perturbs neural differentiation
. JBC. 2021.The isolation of high-quality RNA from endocrine pancreas sections represents a considerable challenge largely due to the high ribonuclease levels. Laser capture microdissection (LCM) of mammalian islets, in association with RNA extraction protocols, has emerged as a feasible approach to characterizing their genetic and proteomic profiles. However, a validated protocol to obtain high-quality RNA from LCM-derived human pancreas specimens that is appropriate for next-generation sequencing analysis is still lacking. In this study, we applied four methods (Picopure extraction kit, Qiazol protocol, Qiazol + Clean-up kit, and RNeasy Microkit + Carrier) to extract RNA from human islets obtained from both non-diabetic individuals and patients with type 2 diabetes who had undergone partial pancreatectomy, as well as handpicked islets from both non-diabetic and diabetic organ donors. The yield and purity of total RNA were determined by 260/280 absorbance using Nanodrop 100 and the RNA integrity number with a bioanalyzer. The results indicated that among the four methods, the RNeasy MicroKit + Carrier (Qiagen) provides the highest yield and purity.