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| Status |
Public on Nov 17, 2015 |
| Title |
Optimizing sgRNA structure to improve CRISPR-Cas9 knockout efficiency |
| Organism |
Homo sapiens |
| Experiment type |
Other
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| Summary |
The current commonly used single-guide RNA (sgRNA) structure has a shortened duplex compared with the native bacterial clustered regularly interspaced short palindromic repeats RNA (crRNA)–transactivating crRNA (tracrRNA) duplex. Here we show that modifying the sgRNA structure by extending the duplex length and mutating the fourth T of the continuous sequence of Ts (which is the pause signal for RNA polymerase III [pol III]) to C or G significantly, and sometimes dramatically, improves knockout efficiency in cells. In addition, the new sgRNA structure also significantly increases the efficiency of more challenging genome-editing procedures, such as gene deletion, which is important for inducing a loss-of-function in non-coding genes.
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| Overall design |
Gene editing efficiency of different sgRNA design in TZM-bl cells
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| Contributor(s) |
Wu H, Dang Y |
| Citation(s) |
26671237 |
| Submission date |
Nov 06, 2015 |
| Last update date |
Jul 26, 2019 |
| Contact name |
Ying Dang |
| E-mail(s) |
ying.dang@ttuhsc.edu
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| Phone |
9152154255
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| Organization name |
TTUHSC
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| Street address |
5001 El Paso Dr
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| City |
El Paso |
| State/province |
TX |
| ZIP/Postal code |
79905 |
| Country |
USA |
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| Platforms (1) |
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| Samples (21)
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| Relations |
| BioProject |
PRJNA301446 |
| SRA |
SRP065896 |
| Supplementary file |
Size |
Download |
File type/resource |
| GSE74766_RAW.tar |
460.0 Kb |
(http)(custom) |
TAR (of XLSX) |
| GSE74766_sgRNAs_sequences.xlsx |
10.8 Kb |
(ftp)(http) |
XLSX |
SRA Run Selector |
| Raw data are available in SRA |
| Processed data provided as supplementary file |
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