| Chapter title |
Application of TALE-Based Approach for Dissecting Functional MicroRNA-302/367 in Cellular Reprogramming
|
|---|---|
| Chapter number | 21 |
| Book title |
MicroRNA Protocols
|
| Published in |
Methods in molecular biology, January 2018
|
| DOI | 10.1007/978-1-4939-7601-0_21 |
| Pubmed ID | |
| Book ISBNs |
978-1-4939-7600-3, 978-1-4939-7601-0
|
| Authors |
Zhonghui Zhang, Wen-Shu Wu, Zhang, Zhonghui, Wu, Wen-Shu |
| Abstract |
MicroRNAs are small 18-24 nt single-stranded noncoding RNA molecules involved in many biological processes, including stemness maintenance and cellular reprogramming. Current methods used in loss-of-function studies of microRNAs have several limitations. Here, we describe a new approach for dissecting miR-302/367 functions by transcription activator-like effectors (TALEs), which are natural effector proteins secreted by Xanthomonas and Ralstonia bacteria. Knockdown of the miR-302/367 cluster uses the Kruppel-associated box repressor domain fused with specific TALEs designed to bind the miR-302/367 cluster promoter. Knockout of the miR-302/367 cluster uses two pairs of TALE nucleases (TALENs) to delete the miR-302/367 cluster in human primary cells. Together, both TALE-based transcriptional repressor and TALENs are two promising approaches for loss-of-function studies of microRNA cluster in human primary cells. |
X Demographics
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| Country | Count | As % |
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| Unknown | 1 | 100% |
Demographic breakdown
| Type | Count | As % |
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| Members of the public | 1 | 100% |
Mendeley readers
Geographical breakdown
| Country | Count | As % |
|---|---|---|
| Unknown | 4 | 100% |
Demographic breakdown
| Readers by professional status | Count | As % |
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| Researcher | 2 | 50% |
| Student > Ph. D. Student | 1 | 25% |
| Unknown | 1 | 25% |
| Readers by discipline | Count | As % |
|---|---|---|
| Biochemistry, Genetics and Molecular Biology | 1 | 25% |
| Agricultural and Biological Sciences | 1 | 25% |
| Unknown | 2 | 50% |