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| Content Provider | World Health Organization (WHO)-Global Index Medicus |
|---|---|
| Author | Du, Dan Qi, Lei S. |
| Description | Author Affiliation: Du D ( Department of Bioengineering, Stanford University, Stanford, California 94305); Qi LS ( Department of Bioengineering, Stanford University, Stanford, California 94305) |
| Abstract | Targeted modulation of transcription is necessary for understanding complex gene networks and has great potential for medical and industrial applications. CRISPR is emerging as a powerful system for targeted genome activation and repression, in addition to its use in genome editing. This protocol describes how to design, construct, and experimentally validate the function of sequence-specific single guide RNAs (sgRNAs) for sequence-specific repression (CRISPRi) or activation (CRISPRa) of transcription in mammalian cells. In this technology, the CRISPR-associated protein Cas9 is catalytically deactivated (dCas9) to provide a general platform for RNA-guided DNA targeting of any locus in the genome. Fusion of dCas9 to effector domains with distinct regulatory functions enables stable and efficient transcriptional repression or activation in mammalian cells. Delivery of multiple sgRNAs further enables activation or repression of multiple genes. By using scaffold RNAs (scRNAs), different effectors can be recruited to different genes for simultaneous activation of some and repression of others. The CRISPRi and CRISPRa methods provide powerful tools for sequence-specific control of gene expression on a genome-wide scale to aid understanding gene functions and for engineering genetic regulatory systems. |
| File Format | HTM / HTML |
| ISSN | 19403402 |
| Issue Number | 1 |
| Volume Number | 2016 |
| e-ISSN | 15596095 |
| Journal | Cold Spring Harbor Protocols |
| Language | English |
| Publisher | Cold Spring Harbor Laboratory Press |
| Publisher Date | 2016-01-04 |
| Publisher Place | United States |
| Access Restriction | One Nation One Subscription (ONOS) |
| Subject Keyword | Discipline Clinical Laboratory Techniques Clustered Regularly Interspaced Short Palindromic Repeats Physiology Databases, Genetic Genome Animals Cells, Cultured Hek293 Cells Humans Rna, Guide Transcription, Genetic Journal Article |
| Content Type | Text |
| Resource Type | Article |
| Subject | Biochemistry, Genetics and Molecular Biology |
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