Long non-coding RNA NEAT1 promotes sarcoma metastasis by regulating RNA splicing pathways

To identify genes that are differentially expressed in lung metastases of sarcoma, we generated KrasLSL-G12D; Trp53Flox/Flox (KP) mice which develop autochthonous sarcomas in the muscle following injection of an adenovirus expressing Cre recombinase. Cre activates expression of mutant KrasG12D and deletes both alleles of Trp53. As each primary sarcoma reached the size of around 300 mm3, the leg bearing the tumor was amputated and the tumor stored for subsequent RNA sequencing. The amputated mice were followed for several months and 6 mice developed 14 lung metastases: two mice developed one lung metastases, two mice developed two lung metastases, and two mice developed four lung metastases respectively. RNA was then extracted from the 6 primary tumors and the 14 matched lung metastases. Therefore, a total of 20 RNA samples were sent for RNA sequencing a total of 20 RNA from the 6 primary tumors and the 14 matched lung metastases were sent for RNA sequencing