Lentivirus-mediated overexpression of miR-122a, a liver specific MicroRNA for gain-of-function study in HCCs.

Diao, Shu. ; Thesis (M.Phil.)--Chinese University of Hong Kong, 2008. ; Includes bibliographical references (leaves 72-73). ; Abstracts in English and Chinese. ; 摘要 --- p.i ; Abstract --- p.ii ; Acknowledgements --- p.iv ; Contents --- p.viii ; Chapter Chapter 1 --- Introduction and Background ; Chapter 1.1 --- General introduction to miRNA --- p.1 ; Chapter 1.1.1 --- The discovery and biogenesis of miRNA --- p.1 ; Chapter 1.1.2 --- The function of miRNA --- p.3 ; Chapter 1.2 --- The liver-specific miRNA : miR-122a --- p.5 ; Chapter 1.2.1 --- Discovery and biogenesis of miR-122a --- p.5 ; Chapter 1.2.2 --- "miR-122a, a liver specific miRNA" --- p.6 ; Chapter 1.2.3 --- miR-122a and Hepatocellular carcinoma (HCC) --- p.6 ; Chapter 1.2.4 --- Therapeutic opportunities and challenges of miR-122a --- p.7 ; Chapter 1.3 --- Techniques and approaches to study miRNAs --- p.8 ; Chapter 1.3.1 --- Discovery of novel miRNA --- p.8 ; Chapter 1.3.2 --- Detection of miRNA --- p.9 ; Chapter 1.3.3 --- Functional study of miRNAs --- p.10 ; Chapter 1.3.4 --- Prediction and validation of miRNA targets --- p.10 ; Chapter 1.4 --- References --- p.12 ; Chapter Chapter 2 --- Materials and methods ; Chapter 2.1 --- Cell culture --- p.18 ; Chapter 2.2 --- Cell transfection --- p.18 ; Chapter 2.3 --- RNA extraction --- p.18 ; Chapter 2.4 --- Plasmid extaction --- p.19 ; Chapter 2.5 --- Competent cell preparation --- p.19 ; Chapter 2.6 --- Bacterial Transformation --- p.20 ; Chapter 2.7 --- Purification of DNA fragments from agarose gel --- p.21 ; Chapter 2.8 --- Genomic DNA extranction from MIHA cell --- p.21 ; Chapter 2.9 --- Real-time RT-PCR analysis --- p.21 ; Chapter 2.10 --- Lenti-vector Construction for miRNA expression --- p.22 ; Chapter 2.11 --- Lentivirus production --- p.22 ; Chapter 2.12 --- Lentiviral vector titering --- p.23 ; Chapter 2.13 --- Bradford protein assay --- p.23 ; Chapter 2.14 --- Western Blot --- p.24 ; Chapter 2.14.1 --- Sample preparation --- p.24 ; Chapter 2.14.2 --- Gel electrophoresis --- p.24 ; Chapter ...