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CRISPR-Cas9 enabled targeted mutagenesis in the thermotolerant methylotrophic yeast Ogataea thermomethanolica

Overview of attention for article published in FEMS Microbiology Letters, April 2018
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Title
CRISPR-Cas9 enabled targeted mutagenesis in the thermotolerant methylotrophic yeast Ogataea thermomethanolica
Published in
FEMS Microbiology Letters, April 2018
DOI 10.1093/femsle/fny105
Pubmed ID
Authors

Chitwadee Phithakrotchanakoon, Aekkachai Puseenam, Sriwan Wongwisansri, Lily Eurwilaichitr, Supawadee Ingsriswang, Sutipa Tanapongpipat, Niran Roongsawang

Abstract

Ogataea thermomethanolica TBRC656 is a thermotolerant methylotrophic yeast suitable for heterologous protein expression at various temperatures. However, the lack of efficient methods for targeted gene mutagenesis limits strain engineering in this yeast. In this study, we applied a CRISPR-Cas9 based-tool for targeted gene mutagenesis in O. thermomethanolica. The putative unfolded protein response (UPR) regulator OtHAC1, and the OtMAL1 (maltase) and OtMAL2 (maltose permease) genes involved with sucrose and maltose utilization were targeted for CRISPR-Cas9 mutagenesis. Plasmids were constructed for integrative and episomal expression of CRISPR-Cas9 elements in O. thermomethanolica in which Cas9 and gRNA are transcribed from the alcohol oxidase (AOX) promoter. The expression of these genome editing elements is controlled by derepression with glycerol and gRNA are flanked by self-cleaving ribozymes. For integrative system, OtHAC1, OtMAL1, and OtMAL2 were disrupted at 63%, 97%, and 93%, respectively. In addition, OtMAL1 was also disrupted with episomal system at 92%. These finding indicate that the CRISPR-Cas9 system described herein are thus applicable for studying gene function and strain engineering in yeast O. thermomethanolica.

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Mendeley readers

The data shown below were compiled from readership statistics for 10 Mendeley readers of this research output. Click here to see the associated Mendeley record.

Geographical breakdown

Country Count As %
Unknown 10 100%

Demographic breakdown

Readers by professional status Count As %
Researcher 4 40%
Student > Ph. D. Student 2 20%
Student > Master 2 20%
Student > Bachelor 1 10%
Unknown 1 10%
Readers by discipline Count As %
Biochemistry, Genetics and Molecular Biology 6 60%
Engineering 2 20%
Agricultural and Biological Sciences 1 10%
Unknown 1 10%

Attention Score in Context

This research output has an Altmetric Attention Score of 1. This is our high-level measure of the quality and quantity of online attention that it has received. This Attention Score, as well as the ranking and number of research outputs shown below, was calculated when the research output was last mentioned on 21 July 2018.
All research outputs
#8,307,898
of 13,255,897 outputs
Outputs from FEMS Microbiology Letters
#2,153
of 2,668 outputs
Outputs of similar age
#160,469
of 268,930 outputs
Outputs of similar age from FEMS Microbiology Letters
#45
of 62 outputs
Altmetric has tracked 13,255,897 research outputs across all sources so far. This one is in the 23rd percentile – i.e., 23% of other outputs scored the same or lower than it.
So far Altmetric has tracked 2,668 research outputs from this source. They receive a mean Attention Score of 4.0. This one is in the 15th percentile – i.e., 15% of its peers scored the same or lower than it.
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We're also able to compare this research output to 62 others from the same source and published within six weeks on either side of this one. This one is in the 16th percentile – i.e., 16% of its contemporaries scored the same or lower than it.