Efficient and rapid fluorescent protein knock-in with universal donors in mouse embryonic stem cells

ABSTRACT: Fluorescent protein (FP) tagging is a key method for observing protein distribution, dynamics, and interaction with other proteins in living cells. However, the typical approach using overexpression of tagged proteins can perturb cell behavior and introduce localization artifacts. To preserve native expression, fluorescent proteins can be inserted directly into endogenous genes. This approach has been widely used in yeast for decades, and more recently in invertebrate model organisms with the advent of CRISPR/Cas9. However, endogenous FP tagging has not been widely used in mammalian cells due to inefficient homology-directed repair. Recently, the CRISPaint system has utilized non-homologous end joining for efficient integration of FP tags into native loci, but it only allows C-terminal kno

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PROVIDER: S-BIAD694 | bioimages |

REPOSITORIES: bioimages