（兰姆）卡尔中U6启动子的鉴定及基因编辑系统的建立

Identification of U6 Promoter and Establishment of Gene-Editing System in (Lamb.) Carr.

作者信息

Xing Jun-Xia, Luo Ao-Jie, Wang Xin-Hao, Ding Qi, Yang Ling, Li Wan-Feng

机构信息

State Key Laboratory of Tree Genetics and Breeding, College of Forestry, Northeast Forestry University, Harbin 150040, China.

State 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, China.

出版信息

Plants (Basel). 2024 Dec 26;14(1):45. doi: 10.3390/plants14010045.

DOI:10.3390/plants14010045

PMID:39795305

原文链接:https://pmc.ncbi.nlm.nih.gov/articles/PMC11722980/

Abstract

This study aimed to establish a CRISPR/Cas9 gene-editing system for (Lamb.) Carr. (Japanese larch). We screened U6 promoters and used them to drive sgRNA expression in the CRISPR/Cas9 gene-editing system. The 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 . Our results demonstrate that the CRISPR/Cas9 system can efficiently edit genes in , with significantly higher editing frequencies observed when sgRNA expression is driven by endogenous promoters compared to the exogenous promoter ProAtU6-26. This work provides technical support for the study of gene functions and genetic improvement.

摘要

本研究旨在建立日本落叶松（Larix kaempferi (Lamb.) Carr.）的CRISPR/Cas9基因编辑系统。我们筛选了U6启动子，并将其用于驱动CRISPR/Cas9基因编辑系统中sgRNA的表达。以日本落叶松胚性愈伤组织作为遗传转化的受体材料，随后分析基因编辑的频率和类型。结果显示转基因材料中出现了各种突变，包括碱基替换和缺失，编辑频率在5%至14.29%之间。总之，我们建立了日本落叶松的CRISPR/Cas9基因编辑系统。我们的结果表明，CRISPR/Cas9系统能够有效地编辑日本落叶松中的基因，与外源启动子ProAtU6-26相比，当由内源U6启动子驱动sgRNA表达时，观察到的编辑频率显著更高。这项工作为日本落叶松基因功能研究和遗传改良提供了技术支持。