Impact of RNA-Guided Technologies for Target Identification and Deconvolution
| Autor: | Qing Xiang, Ralph Garippa, Chun-Hao Huang, Chong Chen, Alexia Hwang, Chi-Chao Chen, Myles Fennell, Raphael Pelossof |
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| Rok vydání: | 2014 |
| Předmět: |
Therapeutic gene modulation
Computational biology Biology Biochemistry Analytical Chemistry Small hairpin RNA RNA interference Animals Drosophila Proteins Humans CRISPR Clustered Regularly Interspaced Short Palindromic Repeats RNA Small Interfering Mammals Genetics Gene knockdown Cas9 RNA-Binding Proteins RNA Genomics High-Throughput Screening Assays Molecular Medicine RNA Interference Functional genomics RNA Guide Kinetoplastida Biotechnology |
| Zdroj: | SLAS Discovery. 19:1327-1337 |
| ISSN: | 2472-5552 |
| Popis: | For well over a decade, RNA interference (RNAi) has provided a powerful tool for investigators to query specific gene targets in an easily modulated loss-of-function setting, both in vitro and in vivo. Hundreds of publications have demonstrated the utility of RNAi in arrayed and pooled-based formats, in a wide variety of cell-based systems, including clonal, stem, transformed, and primary cells. Over the years, there have been significant improvements in the design of target-specific small-interfering RNA (siRNA) and short-hairpin RNA (shRNA), expression vectors, methods for mitigating off-target effects, and accurately interpreting screening results. Recent developments in RNAi technology include the Sensor assay, high-efficiency miR-E shRNAs, improved shRNA virus production with Pasha (DRGC8) knockdown, and assessment of RNAi off-target effects by using the C9-11 method. An exciting addition to the arsenal of RNA-mediated gene modulation is the clustered regularly interspaced short palindromic repeats/Cas9 (CRISPR/Cas) system for genomic editing, allowing for gene functional knockout rather than knockdown. |
| Databáze: | OpenAIRE |
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