Delivery of short hairpin RNAs—triggers of gene silencing—into mouse embryonic stem cells
Christoph Schaniel, Feng Li, Xenia L Schafer, Troy Moore, Ihor R Lemischka
& Patrick J Paddison
Figure 1.Delivery of synthetic shRNA into Nanog-GFP mouse ES cells. (a) Nanog-GFP ES cells. This line was generated by first engineering the mouse BAC RP23-180N22 to contain a gene encoding a Hygromycin-EGFP fusion in place of exon 1 of the Nanog gene and then stably integrating a BsiWI fragment encoding a three-protein fusion of Nanog-Hygromycin-EGFP into CCE mouse ES cells. Scale bar, 100 m. (b) A titration curve of Nanog-GFP ES cells transfected with various amounts of the EGFP_433 shRNA. GFP fluorescence was measured by fluorescence-activated cell sorting (FACS) 72 h after transfection. (c) FACS profiles of Nanog-GFP cells transfected with 20 pmol of EGFP_433 shRNA. The percentage of GFP-expressing cells is shown above the GFP population gate (horizontal bar).
m. (b) A titration curve of Nanog-GFP ES cells transfected with various amounts of the EGFP_433 shRNA. GFP fluorescence was measured by fluorescence-activated cell sorting (FACS) 72 h after transfection. (c) FACS profiles of Nanog-GFP cells transfected with 20 pmol of EGFP_433 shRNA. The percentage of GFP-expressing cells is shown above the GFP population gate (horizontal bar).