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基于软件的查,以寻找有效的sgRNAs在乳球菌乳酸菌中.

Hui Wang1, Lianzhong Ai1, Yongjun Xia1

  • 1Shanghai Engineering Research Center of Food Microbiology, School of Health Science and Engineering, University of Shanghai for Science and Technology, Shanghai, China.

Journal of the science of food and agriculture
|August 30, 2023
PubMed
概括
此摘要是机器生成的。

克里斯普尔基因编辑依赖于单导向RNA (sgRNA) 选择. 这项研究发现,SgRNA预测在诸如乳球菌 (Lactococcus lactis) 等 prokaryotes 并不匹配实验结果,突出需要经验验证.

关键词:
这就是CRISPR/Cas9的作用.乳球球菌 (lactococcus lactis) 是一种乳球菌.基因组工程是基因组工程.在 sgRNA 查中使用 sgRNA 查.

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科学领域:

  • 微生物学 微生物学
  • 分子生物学分子生物学
  • 生物技术是生物技术.

背景情况:

  • 聚类定期间隔的短平行体重复 (CRISPR) 基因编辑依赖于促进子和单导向RNA (sgRNA) 元素.
  • 在CRISPR系统中,sgRNA选择对于精确的基因向和编辑效率至关重要.
  • 在线预测工具可用于根据用户定义的标准预测和排名sgRNA.

研究的目的:

  • 评估在线sgRNA预测工具在原核细胞中的准确性.
  • 为了比较预测的sgRNA效率与Lactococcus lactis的实验结果.
  • 调查真核生物CRISPR发现对 prokaryotic 系统的适用性.

主要方法:

  • 使用CRISPOR在线工具为乳球菌乳糖基因 (ldh和uppp) 设计的sgRNA.
  • 为选择的基因构建淘汰菌株,以评估sgRNA的效率.
  • 将实验编辑效率与软件预测进行了比较.

主要成果:

  • 选择的sgRNAs的实验编辑效率与CRISPOR预测不一致.
  • 在预测和实际sgRNA性能之间观察到显著的差异.
  • 这表明以真核为中心的CRISPR预测模型可能不会直接转化为原核生物.

结论:

  • CRISPR sgRNA预测软件是一种有用的初始查工具,但需要实验验证.
  • 来自真核生物CRISPR研究的既定发现可能不适用于 prokaryotes.
  • 在 prokaryotic 基因编辑中优化 sgRNA 选择,实证测试是必不可少的.