Neutrophil-like HL-60 cells expressing only GFP-tagged β-actin exhibit nearly normal motility.

ABSTRACT: Observations of actin dynamics in living cells using fluorescence microscopy have been foundational in the exploration of the mechanisms underlying cell migration. We used CRISPR/Cas9 gene editing to generate neutrophil-like HL-60 cell lines expressing GFP-β-actin from the endogenous locus (ACTB). In light of many previous reports outlining functional deficiencies of labeled actin, we anticipated that HL-60 cells would only tolerate a monoallelic edit, as biallelic edited cells would produce no normal β-actin. Surprisingly, we recovered viable monoallelic GFP-β-actin cells as well as biallelic edited GFP-β-actin cells, in which one copy of the ACTB gene is silenced and the other contains the GFP tag. Furthermore, the edited cells migrate with similar speeds and persistence as unmodified ce

SUBMITTER: Garner RM 

PROVIDER: S-EPMC7383899 | biostudies-literature | 2020 May

REPOSITORIES: biostudies-literature