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除编辑外,CRISPR/Cas9 用于蛋白质定位:与“dCas9 系统鉴定 Ctf19 在动粒衍生抑制减数分裂重组中的核心作用”一起使用的教育入门

Beyond editing, CRISPR/Cas9 for protein localization: an educational primer for use with "A dCas9-based system identifies a central role for Ctf19 in kinetochore-derived suppression of meiotic recombination".

机构信息

Department of Biology, Hamilton College, Clinton, NY 13323, USA.

Section of Plant Biology, School of Integrative Plant Science, Cornell University, Ithaca, NY 14850, USA.

出版信息

Genetics. 2022 Aug 30;222(1). doi: 10.1093/genetics/iyac109.

Abstract

CRISPR/Cas9 has dramatically changed how we conduct genetic research, providing a tool for precise sequence editing. However, new applications of CRISPR/Cas9 have emerged that do not involve nuclease activity. In the accompanying article "A dCas9-based system identifies a central role for Ctf19 in kinetochore-derived suppression of meiotic recombination," Kuhl et al. utilize a catalytically dead Cas9 to localize proteins at specific genomic locations. The authors seek to understand the role of kinetochore proteins in the suppression of meiotic recombination, a phenomenon that has been observed in centromere regions. By harnessing the power of CRISPR/Cas9 to bind specific genomic sequences, Kuhl et al. localized individual kinetochore proteins to areas of high meiotic recombination and assessed their role in suppression. This primer article provides undergraduate students with background information on chromosomes, meiosis, recombination and CRISPR/Cas9 to support their reading of the Kuhl et al. study. This primer is intended to help students and instructors navigate the study's experimental design, interpret the results, and appreciate the broader scope of meiotic recombination and CRISPR/Cas9. Questions are included to facilitate discussion of the study.

摘要

CRISPR/Cas9 技术极大地改变了我们进行基因研究的方式,为精确的序列编辑提供了一种工具。然而,CRISPR/Cas9 的新应用已经出现,它们不涉及核酸酶活性。在随附的文章“基于 dCas9 的系统确定了 Ctf19 在动粒衍生的减数分裂重组抑制中的核心作用”中,Kuhl 等人利用无催化活性的 Cas9 将蛋白质定位到特定的基因组位置。作者旨在了解动粒蛋白在减数分裂重组抑制中的作用,这种现象在着丝粒区域已经观察到。通过利用 CRISPR/Cas9 结合特定基因组序列的能力,Kuhl 等人将单个动粒蛋白定位到高减数分裂重组区域,并评估它们在抑制中的作用。这篇入门文章为本科生提供了关于染色体、减数分裂、重组和 CRISPR/Cas9 的背景信息,以支持他们阅读 Kuhl 等人的研究。这篇入门文章旨在帮助学生和教师了解该研究的实验设计、解释结果,并欣赏减数分裂重组和 CRISPR/Cas9 的更广泛范围。文中还包含了一些问题,以促进对该研究的讨论。

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Kinetochore Function from the Bottom Up.从底部看动粒功能。
Trends Cell Biol. 2018 Jan;28(1):22-33. doi: 10.1016/j.tcb.2017.09.002. Epub 2017 Oct 3.
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The Kinetochore Receptor for the Cohesin Loading Complex.黏连蛋白装载复合体的动粒受体
Cell. 2017 Sep 21;171(1):72-84.e13. doi: 10.1016/j.cell.2017.08.017.

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