GSM4083953: F5E6_E12.5_Placenta; Mus musculus; RNA-Seq
C57BL6/J females were superovulated and crossed with CAST/EiJ males.  Zygotes were recovered the next day and electroporated with two sgRNAs for the Gab1 RLTR15 ERVK element (200ng/μl each) (Fig S7) and CAS9 protein (500ng/μl).  Embryos were then implanted into NMRI pseudo-pregnant females.  Embryos were dissected on E12.5 and the following tissues were collected: 1) visceral yolk sac/amnion, 2) placental disc with as much decidua removed as possible, 3) embryo, and 4) tail clip for genotyping.  Tissue samples were washed in cold PBS and flash frozen in 50μL of RLT+ buffer (Qiagen).  RNA was extracted from E12.5 yolk sac, placenta and whole embryo using RNeasy Mini Kit (Qiagen) as per the manufacturers' instructions. RNA was treated TURBO DNase (Thermo Fisher Scientific) and quality was assessed with RNA Pico Kit (Agilent) on the Agilent 2100 Bioanalyzer (RIN>8.7 for all samples).  mRNA sequencing libraries were generated using SmartSeq v4 cDNA generation and Nextera XT library preparation, as per manufacturers' instructions.  The quantification of all libraries was done using the High DNA Sensitivity Bioanalyzer 2500 (Agilent) and Illumina library quantification kit (KAPA).  Libraries were sequenced using 125bp paired-end on the Illumina HiSeq 2500 RapidRun, multiplexing 12 samples over two lanes.