Identification of U6 Promoter and Establishment of Gene-Editing System in <i>Larix kaempferi</i> (Lamb.) Carr
This study aimed to establish a CRISPR/Cas9 gene-editing system for <i>Larix kaempferi</i> (Lamb.) Carr. (Japanese larch). We screened <i>L. kaempferi</i> U6 promoters and used them to drive sgRNA expression in the CRISPR/Cas9 gene-editing system. The <i>L. kaempferi<...
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MDPI AG
2024-12-01
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| Series: | Plants |
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| author | Jun-Xia Xing Ao-Jie Luo Xin-Hao Wang Qi Ding Ling Yang Wan-Feng Li |
| author_facet | Jun-Xia Xing Ao-Jie Luo Xin-Hao Wang Qi Ding Ling Yang Wan-Feng Li |
| author_sort | Jun-Xia Xing |
| collection | DOAJ |
| description | This study aimed to establish a CRISPR/Cas9 gene-editing system for <i>Larix kaempferi</i> (Lamb.) Carr. (Japanese larch). We screened <i>L. kaempferi</i> U6 promoters and used them to drive sgRNA expression in the CRISPR/Cas9 gene-editing system. The <i>L. kaempferi</i> embryogenic callus was used as the receptor material for genetic transformation, and the frequency and types of gene editing were then analyzed. The results showed various mutations in the transgenic materials, including base substitutions and deletions, and the editing frequency ranged from 5% to 14.29%. In summary, we established a CRISPR/Cas9 gene-editing system for <i>L. kaempferi</i>. Our results demonstrate that the CRISPR/Cas9 system can efficiently edit genes in <i>L. kaempferi</i>, with significantly higher editing frequencies observed when sgRNA expression is driven by endogenous <i>LaU6</i> promoters compared to the exogenous promoter ProAtU6-26. This work provides technical support for the study of <i>L. kaempferi</i> gene functions and genetic improvement. |
| format | Article |
| id | doaj-art-42cea1e2af2f432485393a13e7c46fa4 |
| institution | Kabale University |
| issn | 2223-7747 |
| language | English |
| publishDate | 2024-12-01 |
| publisher | MDPI AG |
| record_format | Article |
| series | Plants |
| spelling | doaj-art-42cea1e2af2f432485393a13e7c46fa42025-01-10T13:19:35ZengMDPI AGPlants2223-77472024-12-011414510.3390/plants14010045Identification of U6 Promoter and Establishment of Gene-Editing System in <i>Larix kaempferi</i> (Lamb.) CarrJun-Xia Xing0Ao-Jie Luo1Xin-Hao Wang2Qi Ding3Ling Yang4Wan-Feng Li5State Key Laboratory of Tree Genetics and Breeding, College of Forestry, Northeast Forestry University, Harbin 150040, ChinaState Key Laboratory of Tree Genetics and Breeding, Key Laboratory of Tree Breeding and Cultivation of the National Forestry and Grassland Administration, Research Institute of Forestry, Chinese Academy of Forestry, Beijing 100091, ChinaState Key Laboratory of Tree Genetics and Breeding, Key Laboratory of Tree Breeding and Cultivation of the National Forestry and Grassland Administration, Research Institute of Forestry, Chinese Academy of Forestry, Beijing 100091, ChinaLife Science and Technology Center, China National Seed Group Co., Ltd., Wuhan 430073, ChinaState Key Laboratory of Tree Genetics and Breeding, College of Forestry, Northeast Forestry University, Harbin 150040, ChinaState Key Laboratory of Tree Genetics and Breeding, Key Laboratory of Tree Breeding and Cultivation of the National Forestry and Grassland Administration, Research Institute of Forestry, Chinese Academy of Forestry, Beijing 100091, ChinaThis study aimed to establish a CRISPR/Cas9 gene-editing system for <i>Larix kaempferi</i> (Lamb.) Carr. (Japanese larch). We screened <i>L. kaempferi</i> U6 promoters and used them to drive sgRNA expression in the CRISPR/Cas9 gene-editing system. The <i>L. kaempferi</i> embryogenic callus was used as the receptor material for genetic transformation, and the frequency and types of gene editing were then analyzed. The results showed various mutations in the transgenic materials, including base substitutions and deletions, and the editing frequency ranged from 5% to 14.29%. In summary, we established a CRISPR/Cas9 gene-editing system for <i>L. kaempferi</i>. Our results demonstrate that the CRISPR/Cas9 system can efficiently edit genes in <i>L. kaempferi</i>, with significantly higher editing frequencies observed when sgRNA expression is driven by endogenous <i>LaU6</i> promoters compared to the exogenous promoter ProAtU6-26. This work provides technical support for the study of <i>L. kaempferi</i> gene functions and genetic improvement.https://www.mdpi.com/2223-7747/14/1/45CRISPR/Cas9<i>Larix</i><i>SCL6</i>sgRNA |
| spellingShingle | Jun-Xia Xing Ao-Jie Luo Xin-Hao Wang Qi Ding Ling Yang Wan-Feng Li Identification of U6 Promoter and Establishment of Gene-Editing System in <i>Larix kaempferi</i> (Lamb.) Carr Plants CRISPR/Cas9 <i>Larix</i> <i>SCL6</i> sgRNA |
| title | Identification of U6 Promoter and Establishment of Gene-Editing System in <i>Larix kaempferi</i> (Lamb.) Carr |
| title_full | Identification of U6 Promoter and Establishment of Gene-Editing System in <i>Larix kaempferi</i> (Lamb.) Carr |
| title_fullStr | Identification of U6 Promoter and Establishment of Gene-Editing System in <i>Larix kaempferi</i> (Lamb.) Carr |
| title_full_unstemmed | Identification of U6 Promoter and Establishment of Gene-Editing System in <i>Larix kaempferi</i> (Lamb.) Carr |
| title_short | Identification of U6 Promoter and Establishment of Gene-Editing System in <i>Larix kaempferi</i> (Lamb.) Carr |
| title_sort | identification of u6 promoter and establishment of gene editing system in i larix kaempferi i lamb carr |
| topic | CRISPR/Cas9 <i>Larix</i> <i>SCL6</i> sgRNA |
| url | https://www.mdpi.com/2223-7747/14/1/45 |
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