Efficient and flexible tagging of endogenous genes by homology-independent intron targeting

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Figure 3.
Figure 3.

A modified donor allows for easy selection of tagged cells. (A) Schematic of donor constructs without and with a blasticidin resistance (BSD) gene. (B) Enrichment of fluorescence-positive HEK293 mNG21–10 cells tagged with mNG211-BSD(−/+) at CANX intron 14 and CBX1 intron 3 after blasticidin treatment. Data represent mean ± SEM (n = 3). (C) Dot plots of total HEK293 cell populations tagged with mNG211 or with mNG211-BSD(−/+) and selected for 12 d. Plots are colored by density. (D) Confocal microscopy of total cell populations as in C. Images are maximum projections of Z-stacks, and scale bars correspond to 10 µm. (E) Western blot of clonal HAP1 lysates tagged with mClover3 only or mClover3-BSD(−/+) at CANX intron 14, target 1. The values below the anti-CANX blot indicate total levels of the major CANX band (tagged and untagged) relative to levels in wild-type (w.t.) cells.

This Article

  1. Published in Advance June 25, 2019, doi: 10.1101/gr.246413.118 Genome Res. 29: 1322-1328

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