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Fluorescent tagging of endogenous proteins with CRISPR/Cas9 in primary mouse neural stem cells.

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).

SUBMITTER: Morrow CS

PROVIDER: S-EPMC8369070 | biostudies-literature | 2021 Sep

REPOSITORIES: biostudies-literature

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Fluorescent tagging of endogenous proteins with CRISPR/Cas9 in primary mouse neural stem cells.

Morrow Christopher S CS Porter Tiaira J TJ Moore Darcie L DL

STAR protocols 20210814 3

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 <i>in vitro</i>. Here, we describe reagent construction, reagent delivery, and a screening strat ...[more]

PMID: 34430917

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