Methods for analyzing microRNA expression and function during hematopoietic lineage differentiation.
MicroRNAs (miRNAs), an abundant class of approx 22-nucleotide (nt) small RNAs that control gene expression at the posttranscriptional level, may play important roles during normal hematopoiesis and leukemogenesis. This chapter focuses on the methods and strategies for dissecting miRNA function during hematopoietic lineage differentiation. We describe a modified miRNA cloning method and expression analysis approach for determining miRNA expression during hematopoietic lineage differentiation. We illustrate a retroviral vector and a general strategy for the ectopic expression of miRNAs in hemato-poietic stem/progenitor cells. We discuss in vitro and in vivo functional assays that can be used to examine the roles of miRNAs during hematopoietic lineage differentiation. The methods and principles described here should also be applicable to study the roles of miRNAs in the differentiation and function of nonhematopoietic cell types.
Min
Hyeyoung
H
Department of Microbiology and Immunology, Baxter Laboratory of Genetic Pharmacology, Institute for Stem Cell, Cancer and Medicine, Stanford University School of Medicine, Stanford, CA, USA.
Chen
Chang-Zheng
CZ
D000818:Animals; D001402:B-Lymphocytes / Q000166:cytology / Q000378:metabolism; D001854:Bone Marrow Cells / Q000378:metabolism*; D016026:Bone Marrow Transplantation; D002454:Cell Differentiation / Q000235:genetics; D019070:Cell Lineage / Q000235:genetics; D005434:Flow Cytometry; D005786:Gene Expression Regulation; D005822:Genetic Vectors; D006410:Hematopoiesis / Q000235:genetics*; D006412:Hematopoietic Stem Cells / Q000166:cytology / Q000378:metabolism*; D007150:Immunohistochemistry; D066298:In Vitro Techniques; D051379:Mice; D008810:Mice, Inbred C57BL; D008815:Mice, Inbred Strains; D035683:MicroRNAs / Q000737:chemistry / Q000235:genetics / Q000502:physiology*; D012190:Retroviridae / Q000235:genetics; D013601:T-Lymphocytes / Q000166:cytology / Q000378:metabolism
D035683:MicroRNAs