Genome-wide in-locus epitope tagging of Arabidopsis proteins using prime editors.
IF 2.9 3区 生物学Q3 BIOCHEMISTRY & MOLECULAR BIOLOGYBMB ReportsPub Date : 2024-01-01
Cheljong Hong, Jun Hee Han, Gue-Ho Hwang, Sangsu Bae, Pil Joon Seo
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引用次数: 0
Abstract
Prime editors (PEs), which are CRISPR-Cas9 nickase (H840A)-reverse transcriptase fusion proteins programmed with prime editing guide RNAs (pegRNAs), can not only edit bases but also install transversions, insertions, or deletions without both donor DNA and double-strand breaks at the target DNA. As the demand for in-locus tagging is increasing, to reflect gene expression dynamics influenced by endogenous genomic contexts, we demonstrated that PEs can be used to introduce the hemagglutinin (HA) epitope tag to a target gene locus, enabling molecular and biochemical studies using in-locus tagged plants. To promote genome-wide in-locus tagging, we also implemented a publicly available database that designs pegRNAs for in-locus tagging of all the Arabidopsis genes. [BMB Reports 2024; 57(1): 66-70].
初级编辑器（PEs）是CRISPR-Cas9缺口酶（H840A）-逆转录酶融合蛋白，与初级编辑向导RNA（pegRNAs）一起编程，不仅能编辑碱基，还能在靶DNA上安装反转、插入或缺失，而无需供体DNA和双链断裂。为了反映受内源基因组环境影响的基因表达动态，对病灶内标记的需求日益增加，我们证明了 PE 可用于将血凝素（HA）表位标记引入目标基因位点，从而利用病灶内标记的植物进行分子和生化研究。为了促进全基因组的病灶内标记，我们还建立了一个公开可用的数据库，用于设计拟南芥所有基因的病灶内标记的 pegRNA。
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The BMB Reports (BMB Rep, established in 1968) is published at the end of every month by Korean Society for Biochemistry and Molecular Biology. Copyright is reserved by the Society. The journal publishes short articles and mini reviews. We expect that the BMB Reports will deliver the new scientific findings and knowledge to our readers in fast and timely manner.
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