scholarly journals Highly Efficient Targeted Mutagenesis of Drosophila with the CRISPR/Cas9 System

Cell Reports ◽  
2013 ◽  
Vol 4 (1) ◽  
pp. 220-228 ◽  
Author(s):  
Andrew R. Bassett ◽  
Charlotte Tibbit ◽  
Chris P. Ponting ◽  
Ji-Long Liu
2014 ◽  
Vol 4 (1) ◽  
Author(s):  
Akihiro Yasue ◽  
Silvia Naomi Mitsui ◽  
Takahito Watanabe ◽  
Tetsushi Sakuma ◽  
Seiichi Oyadomari ◽  
...  

2018 ◽  
Vol 19 (10) ◽  
pp. 3000 ◽  
Author(s):  
Shouhong Zhu ◽  
Xiuli Yu ◽  
Yanjun Li ◽  
Yuqiang Sun ◽  
Qianhao Zhu ◽  
...  

The clustered regularly interspaced short palindromic repeats/CRISPR-associated protein 9 (CRISPR/Cas9) gene editing system has been shown to be able to induce highly efficient mutagenesis in the targeted DNA of many plants, including cotton, and has become an important tool for investigation of gene function and crop improvement. Here, we developed a simple and easy to operate CRISPR/Cas9 system and demonstrated its high editing efficiency in cotton by targeting-ALARP, a gene encoding alanine-rich protein that is preferentially expressed in cotton fibers. Based on sequence analysis of the target site in the 10 transgenic cottons containing CRISPR/Cas9, we found that the mutation frequencies of GhALARP-A and GhALARP-D target sites were 71.4–100% and 92.9–100%, respectively. The most common editing event was deletion, but deletion together with large insertion was also observed. Mosaic mutation editing events were detected in most transgenic plants. No off-target mutation event was detected in any the 15 predicted sites analyzed. This study provided mutants for further study of the function of GhALARP in cotton fiber development. Our results further demonstrated the feasibility of use of CRISPR/Cas9 as a targeted mutagenesis tool in cotton, and provided an efficient tool for targeted mutagenesis and functional genomics in cotton.


Development ◽  
2014 ◽  
Vol 141 (10) ◽  
pp. 2165-2171 ◽  
Author(s):  
G. P. Flowers ◽  
A. T. Timberlake ◽  
K. C. Mclean ◽  
J. R. Monaghan ◽  
C. M. Crews

Cell Reports ◽  
2014 ◽  
Vol 6 (6) ◽  
pp. 1178-1179 ◽  
Author(s):  
Andrew R. Bassett ◽  
Charlotte Tibbit ◽  
Chris P. Ponting ◽  
Ji-Long Liu

2019 ◽  
Vol 17 (11) ◽  
pp. 2032-2034 ◽  
Author(s):  
Toshihisa Nomura ◽  
Komaki Inoue ◽  
Yukiko Uehara‐Yamaguchi ◽  
Koji Yamada ◽  
Osamu Iwata ◽  
...  

2014 ◽  
Vol 49 ◽  
pp. 35-42 ◽  
Author(s):  
Yuanyuan Liu ◽  
Sanyuan Ma ◽  
Xiaogang Wang ◽  
Jiasong Chang ◽  
Jie Gao ◽  
...  

Genetics ◽  
2013 ◽  
Vol 195 (3) ◽  
pp. 703-713 ◽  
Author(s):  
Sudeepta Kumar Panda ◽  
Benedikt Wefers ◽  
Oskar Ortiz ◽  
Thomas Floss ◽  
Bettina Schmid ◽  
...  

Author(s):  
Trevor Weiss ◽  
Chunfang Wang ◽  
Xiaojun Kang ◽  
Hui Zhao ◽  
Maria Elena Gamo ◽  
...  

AbstractIn recent years, Setaria viridis has been developed as a model plant to better understand the C4 photosynthetic pathway in major crops. With the increasing availability of genomic resources for S. viridis research, highly efficient genome editing technologies are needed to create genetic variation resources for functional genomics. Here, we developed a protoplast assay to rapidly optimize the multiplexed CRISPR/Cas9 system in S. viridis. Targeted mutagenesis efficiency was further improved by an average of 1.4-fold with the exonuclease, Trex2. Distinctive mutation profiles were found in the Cas9_Trex2 samples with 94% of deletions larger than 10bp, and less than 1% of mutations being insertions. Further analyses indicated that 52.2% of deletions induced by Cas9_Trex2, as opposed to 3.5% by Cas9 alone, were repaired through microhomology-mediated end joining (MMEJ) rather than the canonical NHEJ DNA repair pathway. Combined with the robust agrobacterium-mediated transformation method with more than 90% efficiency, the multiplex CRISPR/Cas9_Trex2 system was demonstrated to induce targeted mutations in two tightly linked genes, svDrm1a and svDrm1b, at the frequency ranging from 73% to 100% in T0 plants. These mutations were transmitted to at least 60% of the transgene-free T1 plants with 33% of them containing bi-allelic or homozygous mutations in both genes. This highly efficient multiplex CRISPR/Cas9_Trex2 system makes it possible to create a large mutant resource for S. viridis in a rapid and high throughput manner, and has the potential to be widely applicable in achieving more predictable MMEJ-mediated mutations in many plant species.


2020 ◽  
Vol 56 (27) ◽  
pp. 3851-3854 ◽  
Author(s):  
Xiaomin Chai ◽  
Hai-Hua Huang ◽  
Huiping Liu ◽  
Zhuofeng Ke ◽  
Wen-Wen Yong ◽  
...  

A Co-based complex displayed the highest photocatalytic performance for CO2 to CO conversion in aqueous media.


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