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journal.pone.0276657.pdf | 3 MB | Adobe PDF | 見る/開く |
タイトル: | Evaluation of a library of loxP variants with a wide range of recombination efficiencies by Cre |
著者: | Yamauchi, Yuji Matsukura, Hidenori Motone, Keisuke Ueda, Mitsuyoshi Aoki, Wataru |
著者名の別形: | 山内, 悠至 松倉, 秀典 植田, 充美 青木, 航 |
キーワード: | Saccharomyces cerevisiae Mutagenesis Gene expression Yeast Polymerase chain reaction Recombinases Substitution mutation Nucleotides |
発行日: | 21-Oct-2022 |
出版者: | Public Library of Science (PLoS) |
誌名: | PLOS ONE |
巻: | 17 |
号: | 10 |
論文番号: | e0276657 |
抄録: | Sparse labeling of individual cells is an important approach in neuroscience and many other fields of research. Various methods have been developed to sparsely label only a small population of cells; however, there is no simple and reproducible strategy for managing the probability of sparse labeling at desired levels. Here, we aimed to develop a novel methodology based on the Cre-lox system to regulate sparseness at desired levels, and we purely analyzed cleavage efficiencies of loxP mutants by Cre. We hypothesized that mutations in the loxP sequence reduce the recognition efficiency by Cre, which enables the regulation of the sparseness level of gene expression. In this research, we mutagenized the loxP sequence and analyzed a library of loxP variants. We evaluated more than 1000 mutant loxP sequences, including mutants with reduced excision efficiencies by Cre ranging from 0.51% to 59%. This result suggests that these mutant loxP sequences can be useful in regulating the sparseness of genetic labeling at desired levels. |
著作権等: | © 2022 Yamauchi et al. This is an open access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited. |
URI: | http://hdl.handle.net/2433/285966 |
DOI(出版社版): | 10.1371/journal.pone.0276657 |
PubMed ID: | 36269789 |
出現コレクション: | 学術雑誌掲載論文等 |
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