| Chapter title |
Downregulation of human Cdc6 protein using a lentivirus RNA interference expression vector.
|
|---|---|
| Chapter number | -1597450944 |
| Book title |
MicroRNA Protocols
|
| Published in |
Methods in molecular biology, December 2005
|
| DOI | 10.1385/1-59745-123-1:287 |
| Pubmed ID | |
| Book ISBNs |
978-1-58829-581-1, 978-1-59745-123-9
|
| Authors |
Luo F, Yee JK, Huang SH, Wu LT, Jong AY, Feng Luo, Jiing-Kuan Yee, Sheng-He Huang, Ling-Tao Wu, Ambrose Y. Jong |
| Abstract |
Eukaryotic CDC6 gene function is required for the initiation of DNA replication and is a key regulatory protein during cell cycle progression. The human CDC6 gene is not expressed in most normal tissues, in contrast with its marked expression in proliferating cancer cells. An effective way to explore the gene functions of CDC6 is to knock-down the CDC6 messenger RNA (mRNA) and examine the phenotypic consequences. In this chapter, we describe the construction of a lentivirus vector to express a CDC6 DNA segment. The transcript is able to fold by itself because the sense and antisense regions are complementary. There is a 9-nucleotide (nt) loop region allowing for the short hairpin RNA (shRNA) to form. Cellular ribonucleases process the shRNA into a functional short interfering RNA (siRNA). Down-regulation of Cdc6 protein is confirmed by Western blots. |
X Demographics
Geographical breakdown
| Country | Count | As % |
|---|---|---|
| Unknown | 1 | 100% |
Demographic breakdown
| Type | Count | As % |
|---|---|---|
| Members of the public | 1 | 100% |
Mendeley readers
Geographical breakdown
| Country | Count | As % |
|---|---|---|
| United States | 2 | 8% |
| Unknown | 23 | 92% |
Demographic breakdown
| Readers by professional status | Count | As % |
|---|---|---|
| Researcher | 11 | 44% |
| Student > Ph. D. Student | 5 | 20% |
| Professor | 3 | 12% |
| Other | 1 | 4% |
| Lecturer | 1 | 4% |
| Other | 2 | 8% |
| Unknown | 2 | 8% |
| Readers by discipline | Count | As % |
|---|---|---|
| Agricultural and Biological Sciences | 12 | 48% |
| Biochemistry, Genetics and Molecular Biology | 3 | 12% |
| Medicine and Dentistry | 2 | 8% |
| Computer Science | 2 | 8% |
| Pharmacology, Toxicology and Pharmaceutical Science | 1 | 4% |
| Other | 2 | 8% |
| Unknown | 3 | 12% |