当前位置: X-MOL 学术Plant Mol. Biol. › 论文详情
Our official English website, www.x-mol.net, welcomes your feedback! (Note: you will need to create a separate account there.)
Gene editing in plants: assessing the variables through a simplified case study.
Plant Molecular Biology ( IF 3.9 ) Pub Date : 2020-02-10 , DOI: 10.1007/s11103-020-00976-2
Jay Shockey 1
Affiliation  

Multiple variables that control the relative levels of successful heritable plant genome editing were addressed using simple case studies in Arabidopsis thaliana. The recent advent of genome editing technologies (especially CRISPR, Clustered Regularly Interspaced Short Palindromic Repeats) has revolutionized various fields of scientific research. The process is much more specific than previous mutagenic processes and allows for targeting of nearly any gene of interest for the creation of loss-of-function mutations and many other types of editing, including gene-replacement and gene activation. However, not all CRISPR construct designs are successful, due to several factors, including differences in the strength and cell- or tissue-type specificity of the regulatory elements used to express the Cas9 (CRISPR Associated protein 9) DNA nuclease and single guide RNA components, and differences in the relative editing efficiency at different target areas within a given gene. Here we compare the levels of editing created in Arabidopsis thaliana by CRISPR constructs containing either different promoters, or altered target sites with varied levels of guanine-cytosine base content. Additionally, nuclease activity at sites targeted by imperfectly matched single guide RNAs was observed, suggesting that while the primary goal of most CRISPR construct designs is to achieve rapid, robust, heritable gene editing, the formation of unintended mutations at other genomic loci must be carefully monitored.

中文翻译:

植物中的基因编辑:通过简化的案例研究评估变量。

使用拟南芥中的简单案例研究解决了控制成功的可遗传植物基因组编辑的相对水平的多个变量。最近出现的基因组编辑技术(尤其是 CRISPR,成簇的规则间隔短回文重复序列)已经彻底改变了科学研究的各个领域。该过程比以前的诱变过程更具体,并且允许靶向几乎任何感兴趣的基因,以创建功能丧失突变和许多其他类型的编辑,包括基因替换和基因激活。然而,并非所有的 CRISPR 构建体设计都是成功的,这有几个因素,包括用于表达 Cas9(CRISPR 相关蛋白 9)DNA 核酸酶和单向导 RNA 成分的调节元件的强度和细胞或组织类型特异性的差异,以及给定范围内不同目标区域的相对编辑效率的差异基因。在这里,我们比较了 CRISPR 构建体在拟南芥中创建的编辑水平,该构建体包含不同的启动子,或具有不同鸟嘌呤-胞嘧啶碱基含量水平的改变的靶位点。此外,观察到不完全匹配的单向导 RNA 靶向位点的核酸酶活性,这表明虽然大多数 CRISPR 构建体设计的主要目标是实现快速、稳健、可遗传的基因编辑,但必须小心地在其他基因组位点形成意外突变被监控。以及给定基因内不同目标区域的相对编辑效率的差异。在这里,我们比较了 CRISPR 构建体在拟南芥中创建的编辑水平,该构建体包含不同的启动子,或具有不同鸟嘌呤-胞嘧啶碱基含量水平的改变的靶位点。此外,观察到不完全匹配的单向导 RNA 靶向位点的核酸酶活性,这表明虽然大多数 CRISPR 构建体设计的主要目标是实现快速、稳健、可遗传的基因编辑,但必须小心地在其他基因组位点形成意外突变被监控。以及给定基因内不同目标区域的相对编辑效率的差异。在这里,我们比较了 CRISPR 构建体在拟南芥中创建的编辑水平,该构建体包含不同的启动子,或具有不同鸟嘌呤-胞嘧啶碱基含量水平的改变的靶位点。此外,观察到不完全匹配的单向导 RNA 靶向位点的核酸酶活性,这表明虽然大多数 CRISPR 构建体设计的主要目标是实现快速、稳健、可遗传的基因编辑,但必须小心地在其他基因组位点形成意外突变被监控。或改变具有不同水平的鸟嘌呤-胞嘧啶碱基含量的靶位点。此外,观察到不完全匹配的单向导 RNA 靶向位点的核酸酶活性,这表明虽然大多数 CRISPR 构建体设计的主要目标是实现快速、稳健、可遗传的基因编辑,但必须小心地在其他基因组位点形成意外突变被监控。或改变具有不同水平的鸟嘌呤-胞嘧啶碱基含量的靶位点。此外,观察到不完全匹配的单向导 RNA 靶向位点的核酸酶活性,这表明虽然大多数 CRISPR 构建体设计的主要目标是实现快速、稳健、可遗传的基因编辑,但必须小心地在其他基因组位点形成意外突变被监控。
更新日期:2020-04-22
down
wechat
bug