{Reference Type}: Journal Article
{Title}: Insulin secretion assays in an engineered MIN6 cell line.
{Author}: Yang L;Chen W;
{Journal}: MethodsX
{Volume}: 10
{Issue}: 0
{Year}: 2023
暂无{DOI}: 10.1016/j.mex.2023.102029
{Abstract}: Insulin secretion from pancreatic beta cells is crucial for maintaining glucose homeostasis. The murine insulinoma derived MIN6 cell line is commonly used as a model for insulin secretion studies. However, its glucose responsiveness wanes with passaging, and insulin secretion is traditionally measured by expensive and time-consuming RIA or ELISA. We have developed a MIN6 subclone (MIN6-6) that allows for high throughput assay of insulin secretion in both population and single cells. In addition, MIN6-6 also expresses Cas9, permitting genome wide CRISPR screen of insulin secretion using a pooled sgRNA library. Here we provide methods for assaying insulin secretion both in bulk and in single cells in MIN6-6 cells, as well as for CRISPR screen of insulin secretion.•A highly glucose responsive beta cell reporter line (MIN6-6) with multiple engineered functionalities.•Allows for CRISPR/Cas9 mutagenesis, quantification of bulk insulin secretion by a straightforward nanoLuc assay and visualization of intracellular insulin granules.•Allows for en masse quantification of insulin granule exocytosis in individual cells under multiple conditions.