scholarly journals Highly efficient targeted mutagenesis in one-cell mouse embryos mediated by the TALEN and CRISPR/Cas systems

2014 ◽  
Vol 4 (1) ◽  
Author(s):  
Akihiro Yasue ◽  
Silvia Naomi Mitsui ◽  
Takahito Watanabe ◽  
Tetsushi Sakuma ◽  
Seiichi Oyadomari ◽  
...  
Keyword(s):  
Mouse Embryos ◽  
Targeted Mutagenesis ◽  
Highly Efficient

scholarly journals Highly Efficient Targeted Gene Editing in Upland Cotton Using the CRISPR/Cas9 System

2018 ◽  
Vol 19 (10) ◽  
pp. 3000 ◽  
Author(s):  
Shouhong Zhu ◽  
Xiuli Yu ◽  
Yanjun Li ◽  
Yuqiang Sun ◽  
Qianhao Zhu ◽  
...  
Keyword(s):  
Gene Editing ◽  
Crop Improvement ◽  
Targeted Mutagenesis ◽  
Cotton Fibers ◽  
Efficient Tool ◽  
Gene Encoding ◽  
Editing Event ◽  
Highly Efficient ◽  
Target Sites ◽  
Targeted Gene Editing

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.

scholarly journals 337. CRISPR/Cas9 Mediated Highly Efficient Genome Engineering in Mouse Embryos

2015 ◽  
Vol 23 ◽  
pp. S135
Author(s):  
Khurshida Begum ◽  
Bert W. O'Malley ◽  
Francesco J. DeMayo ◽  
Paul Overbeek
Keyword(s):  
Genome Engineering ◽  
Mouse Embryos ◽  
Highly Efficient

scholarly journals Prime editing primarily induces undesired outcomes in mice

2020 ◽  
Author(s):  
Tomomi Aida ◽  
Jonathan J. Wilde ◽  
Lixin Yang ◽  
Yuanyuan Hou ◽  
Mengqi Li ◽  
...  
Keyword(s):  
Genome Editing ◽  
High Frequency ◽  
Mouse Embryos ◽  
Biomedical Science ◽  
New Approach ◽  
Chemically Modified ◽  
Highly Efficient ◽  
Large Deletions

SummaryGenome editing has transformed biomedical science, but is still unpredictable and often induces undesired outcomes. Prime editing (PE) is a promising new approach due to its proposed flexibility and ability to avoid unwanted indels. Here, we show highly efficient PE-mediated genome editing in mammalian zygotes. Utilizing chemically modified guideRNAs, PE efficiently introduced 10 targeted modifications including substitutions, deletions, and insertions across 6 genes in mouse embryos. However, we unexpectedly observed a high frequency of undesired outcomes such as large deletions and found that these occurred more often than pure intended edits across all of the edits/genes. We show that undesired outcomes result from the double-nicking PE3 strategy, but that omission of the second nick largely ablates PE function. However, sequential double-nicking with PE3b, which is only applicable to a fraction of edits, eliminated undesired outcomes. Overall, our findings demonstrate the promising potential of PE for predictable, flexible, and highly efficient in vivo genome editing, but highlight the need for improved variations of PE before it is ready for widespread use.

scholarly journals Highly efficient targeted mutagenesis in axolotl using Cas9 RNA-guided nuclease

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
Keyword(s):  
Targeted Mutagenesis ◽  
Highly Efficient

Highly efficient RNA-guided base editing in mouse embryos

2017 ◽  
Vol 35 (5) ◽  
pp. 435-437 ◽  
Author(s):  
Kyoungmi Kim ◽  
Seuk-Min Ryu ◽  
Sang-Tae Kim ◽  
Gayoung Baek ◽  
Daesik Kim ◽  
...  
Keyword(s):  
Mouse Embryos ◽  
Base Editing ◽  
Highly Efficient

scholarly journals Electroporation enables the efficient mRNA delivery into the mouse zygotes and facilitates CRISPR/Cas9-based genome editing

2015 ◽  
Vol 5 (1) ◽  
Author(s):  
Masakazu Hashimoto ◽  
Tatsuya Takemoto
Keyword(s):  
Genetic Analysis ◽  
Genome Editing ◽  
High Throughput ◽  
Large Scale ◽  
Mouse Embryos ◽  
Mutant Mice ◽  
Highly Efficient ◽  
Mouse Zygotes ◽  
Time Required

Abstract Recent use of the CRISPR/Cas9 system has dramatically reduced the time required to produce mutant mice, but the involvement of a time-consuming microinjection step still hampers its application for high-throughput genetic analysis. Here we developed a simple, highly efficient and large-scale genome editing method, in which the RNAs for the CRISPR/Cas9 system are electroporated into zygotes rather than microinjected. We used this method to perform single-stranded oligodeoxynucleotide (ssODN)-mediated knock-in in mouse embryos. This method facilitates large-scale genetic analysis in the mouse.

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

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