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促进马铃薯中的基因编辑:Solanum tuberosum L. cv. Desiree 基因组的单核苷酸多态性 (SNP) 图谱。

Facilitating gene editing in potato: a Single-Nucleotide Polymorphism (SNP) map of the Solanum tuberosum L. cv. Desiree genome.

机构信息

Univ. Lille, CNRS, UMR8576 - UGSF - Unité de Glycobiologie Structurale et Fonctionnelle, F-59000, Lille, France.

Univ. Lille, CNRS, USR 3290 - MSAP - Miniaturisation pour la Synthèse l'Analyse et la Protéomique, F-59000, Lille, France.

出版信息

Sci Rep. 2020 Feb 6;10(1):2045. doi: 10.1038/s41598-020-58985-6.

Abstract

Genome editing is a powerful tool for plant functional genomics allowing for multiallelic targeted mutagenesis. The recent development of Clustered Regularly Interspaced Short Palindromic Repeats (CRISPR)/CRISPR associated protein 9 (Cas9) systems for gene editing in plants allows for simple, cost-effective introduction of site-specific double-stranded DNA breaks. The nuclear genomes of a homozygous doubled-monoploid potato clone (DM) and a heterozygous diploid clone (RH) have been sequenced in 2011. However, common potato cultivars display a highly heterozygous autotetraploid genome thus complicating target design for tetra-allelic gene editing. Here, we report on the SNP physical map of the widely used Solanum tuberosum L. cv. Desiree and on the position of the diverse indels providing an essential tool for target design in genome editing approaches. We used this tool for designing a specific gRNA and successfully knocking-out a newly discovered starch synthase gene (SS6) in potato. Resequencing data are publicly available at the Sequence Read Archive of the NCBI (accession number: PRJNA507597) and will represent a valuable resource for functional genomic studies of various metabolic pathways, cell and plant physiology as well as high-throughput reverse genetics in potato.

摘要

基因组编辑是一种强大的植物功能基因组学工具,可实现多等位基因靶向诱变。最近开发的用于植物基因编辑的成簇规律间隔短回文重复(CRISPR)/CRISPR 相关蛋白 9(Cas9)系统允许简单、经济高效地引入靶向双链 DNA 断裂。2011 年对纯合双倍单倍体马铃薯克隆(DM)和杂合二倍体克隆(RH)的核基因组进行了测序。然而,常见的马铃薯品种显示出高度杂合的同源四倍体基因组,因此使针对四等位基因基因编辑的目标设计复杂化。在这里,我们报告了广泛使用的 Solanum tuberosum L. cv. Desiree 的 SNP 物理图谱和不同插入缺失的位置,为基因组编辑方法中的目标设计提供了必要的工具。我们使用该工具设计了一个特定的 gRNA,并成功敲除了马铃薯中新发现的淀粉合酶基因(SS6)。重测序数据可在 NCBI 的序列读取档案库(访问号:PRJNA507597)中公开获取,将成为各种代谢途径、细胞和植物生理学以及马铃薯高通量反向遗传学的功能基因组学研究的有价值资源。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/1e4c/7005264/388bd0ae5116/41598_2020_58985_Fig1_HTML.jpg

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