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. 2012 May 7;21(16):2981–2986. doi: 10.1089/scd.2012.0065

FIG. 1.

FIG. 1.

Identification of Rat Rosa26 locus and gene targeting of rat pluripotent stem cells by inserting a cassette containing tdTomato into Rosa26 locus. (A) The RT-PCR analysis of rat Rosa26 noncoding RNA in rat ESCs. The primer positions are schematically shown by arrowheads under the predicted noncoding RNA transcript. (B) Quantitative real-time PCR of Rosa26 noncoding RNA in rat ESCs and adult tissues relative to β-actin. Error bars are mean±SD (n=2). (C) Structure of targeting vectors and a schematic of knock-in genome of rat Rosa26 locus on chromosome 4. Letters on the locus indicate selected restriction enzyme sites. A, ApaI; E, EcoR1; H, HindIII; N, NheI; Xb, XbaI; Xh, XhoI. (D) Result of the Southern blot analysis using ApaI-digested genomic DNA hybridized with 5′ probe schematically shown in (A). (E) Photomicrograph of Rosa26-tdTomato knock-in rat ESCs generated from the BLK2i-1 line (BLK-RT2). These ESCs express tdTomato ubiquitously. ESCs, embryonic stem cells; RT-PCR, reverse transcription–polymerase chain reaction.