A Facile Method to Append a Bio-ID Tag to Endogenous Mutant Kras Alleles.
| Autor: | Budagyan K; Drexel University College of Medicine, Philadelphia, PA, USA.; Fox Chase Cancer Center, Philadelphia, PA, USA., Cannon AC; Drexel University College of Medicine, Philadelphia, PA, USA.; Fox Chase Cancer Center, Philadelphia, PA, USA., Chernoff J; Fox Chase Cancer Center, Philadelphia, PA, USA. Jonathan.Chernoff@fccc.edu. |
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| Jazyk: | angličtina |
| Zdroj: | Methods in molecular biology (Clifton, N.J.) [Methods Mol Biol] 2024; Vol. 2797, pp. 351-362. |
| DOI: | 10.1007/978-1-0716-3822-4_25 |
| Abstrakt: | KRAS mutations occur in approximately ~50% of colorectal cancers (CRCs) and are associated with poor prognosis and resistance to therapy. While these most common mutations found at amino acids G12, G13, Q61, and A146 have long been considered oncogenic drivers of CRC, emerging clinical data suggest that each mutation may possess different biological functions, resulting in varying consequences in oncogenesis. Currently, the mechanistic underpinnings associated with each allelic variation remain unclear. Elucidating the unique effectors of each KRAS mutant could both increase the understanding of KRAS biology and provide a basis for allele-specific therapeutic opportunities. Biotinylation identification (BioID) is a method to label and identify proteins located in proximity of a protein of interest. These proteins are captured through the strong interaction between the biotin label and streptavidin bead and subsequently identified by mass spectrometry. Here, we developed a protocol using CRISPR-mediated gene editing to generate endogenous BioID (© 2024. The Author(s), under exclusive license to Springer Science+Business Media, LLC, part of Springer Nature.) |
| Databáze: | MEDLINE |
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