MicroRNA cloning from cells of the immune system.
MicroRNAs have emerged as - important posttranscriptional regulators of gene expression. Small RNA cloning is a powerful method to identify new microRNAs (miRNAs) and to profile miRNA expression. In addition, it reveals end heterogeneity that may be important in miRNA function. Here, we describe a protocol that is optimized to clone small RNAs from limited amounts of starting material. This is often the case for studying miRNAs in a highly purified population of immune cells or other primary cell types with limited numbers. The small RNAs cloned with this protocol will have a 5'-PO(4) and 3'-OH group, typical features of miRNAs, so majority of the cloned small RNAs will be miRNAs.
Wu
Haoquan
H
Department of Biomedical Sciences, Center of Excellence in Infectious Disease Research, Paul L. Foster School of Medicine, Texas Tech University Health Sciences Center, El Paso, TX, USA. haoquan.wu@ttuhsc.edu
Neilson
Joel
J
Manjunath
N
N
D000818:Animals; D003001:Cloning, Molecular / Q000379:methods*; D006801:Humans; D007107:Immune System / Q000033:anatomy & histology* / Q000502:physiology*; D035683:MicroRNAs / Q000235:genetics*; D021141:Nucleic Acid Amplification Techniques / Q000295:instrumentation / Q000379:methods; D009693:Nucleic Acid Hybridization / Q000379:methods; D016133:Polymerase Chain Reaction / Q000295:instrumentation / Q000379:methods
D035683:MicroRNAs