Genome-wide analysis of miRNA-mRNA interactions in the marrow microenvironment (HTS)

We report genome-wide analysis of miRNA-mRNA interactions in the hematopoietic marrow microenvironment (ME) by employing the biochemical technique. High-throughput sequencing of RNA isolated by crosslinking immunoprecipitation or HITS-CLIP. Specifically, we analyzed 3 kinds of stromal cells (two human stromal cell lines named HS5 and HS27a and primary mesenchymal stromal cells (MSC) from normal donors) and two types of endothelial cells (TrBMEC, a human bone marrow endothelial cell line) and HUVEC (Human Umbilical Vein Endothelial Cells). Immune precipitation (IP) of Argonaut proteins (AGO1-4) was performed by anti-AGO monoclonal antibody 2A8. IP was performed on cells cross-linked by short-wave ultraviolet radiation (UV) 365 nM. Protocol was modified from as originally published by Chi et al Nature. 2009 Jul 23;460(7254):479-86.