Fluorescent tagging of endogenous proteins with CRISPR/Cas9 in primary mouse neural stem cells
Year: 2021
Type: article
Abstract: Although exogenous overexpression of a protein fused to a fluorescent tag can provide insight for the protein’s function, it also can produce artifacts attributed to its upregulation and may not fully report the endogenous regulation of the protein of interest. To circumvent these issues, we adapted a protocol to label endogenous proteins with fluorescent tags in primary adult mouse neural stem cells in vitro. Here, we describe reagent construction, reagent delivery, and a screening strategy to isolate edited cells. For complete details on the use and execution of this protocol, please refer to Morrow et al. (2020).
Source: STAR Protocols
Institution: University of Wisconsin–Madison
Language: English
Subfield: Molecular Biology
Domain: Life Sciences
Open Access status: gold
Estimated APC paid: $1,500
Funders: Alfred P. Sloan Foundation, American Federation for Aging Research, Wisconsin Alumni Research Foundation, American Foundation for Aging Research, University of Wisconsin-Madison, Greater Milwaukee Foundation, Vallee Foundation, National Institutes of Health, National Institute on Aging, NIH Office of the Director, National Cancer Institute, National Institute of General Medical Sciences, National Institute of Neurological Disorders and Stroke, National Center for Research Resources
Awards: Multi-color Benchtop Flow Cytometer, G5827117300, G6049301734, The mechanisms and functional role of the asymmetric segregation of cellular cargoes in stem cells, Training Program In Molecular AND Cellular Pharmacology, G7741799378, G8857535580, G9000352744, UW Comprehensive Cancer Center Support, Biology of Aging and Age-Related Diseases
OpenAlex ID: W3196063683