Pluto Biosciences, Inc

GSE116628: RNA-seq of Wnt7b overexpressing bone

Bulk RNA sequencing

We report the application of RNA sequencing technology for high-throughput profiling of gene expression in Wnt7b overexpressing bone and wide-type bones.To increase Wnt7b in bone we crossed 9.6kb DMP1-Cre mice and R26-Wnt7b mice. 9.6kb DMP1-Cre; R26-Wnt7b mice were set as OE mice meanwhile their littermates of R26-Wnt7b were controls (Ctrl). Owing to heterozygotes and homozygotes of OE mice didnt show any different phenotypes all OE mice used in this study were homozygotes. Total mRNA of femur was collected from 8-week-old OE and Ctrl mice respectively. Fresh femur samples without soft tissues were rapidly collected within 2 min after euthanasia. And then samples were immediately transported into a sterile 10cm culture dish containing cold PBS, followed with articular cartilage removal via sterile lancets within 5 min, being flushed 3 times using sterile cold PBS to remove marrow cells and being chopped into 2~5mm fragments using sterile scissors. About 200mg bone fragments were collected into a sterile mortal containing 1ml liquid nitrogen and then were crushed and homogenized using a sterile pestle within 5 min, followed by a sterile scalpel until the tissue was adequately homogenized. Homogenized samples were collected into a 2-ml sterile tube, followed with 1 ml TRIzol (Invitrogen, CA, USA) solution added. Then all the extraction procedures were conducted according to manufactures instructions for RNA extraction from homogenized tissues. After quality checks they were subjected to the high-throughput RNA-sequencing. Every group consists of 3 replicates which were from 3 different mice. SOURCE: Fanyuan Yu (fanyuan_yu@outlook.com) - Craniofacial Repair and Regeneration State Key Laboratory of Oral Diseases, West China Hospital of Stomatology

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