Epigenetic state and gene expression remain stable after CRISPR/Cas-mediated chromosomal inversions

IF 8.1 1区 生物学 Q1 PLANT SCIENCES New Phytologist Pub Date : 2025-01-29 DOI:10.1111/nph.20403
Solmaz Khosravi, Rebecca Hinrichs, Michelle Rönspies, Reza Haghi, Holger Puchta, Andreas Houben
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CRISPR/ cas介导的染色体倒位后,表观遗传状态和基因表达保持稳定
DNA序列的染色体位置、染色质的表观遗传状态以及基因活性之间存在着普遍的相关性(Grewal &;莫札,2003;刘等人,2016)。染色体臂是富含常染色质的,而在许多物种中着丝粒和周围着丝粒区域是异色的(Roudier等,2009)。常染色质是染色质的去浓缩部分,主要含有活性基因(Strahl et al., 1999)。相比之下,异染色质,即浓缩染色质部分,在基因和基因活性方面都很差(Fischer et al., 2006;刘等人,2016)。染色质状态的形成和维持受DNA甲基化和翻译后组蛋白修饰的表观遗传调控。异染色质富含高甲基化的DNA和二甲基化的组蛋白H3K9 (H3K9me2) (Soppe et al., 2002)。相比之下,常染色质与三甲基化的H3K4 (H3K4me3)和较少的DNA c -甲基化相关联。位置效应斑纹(PEV),发现于果蝇(Gowen &;Gay, 1934)和人类(Finelli et al., 2012),以及在出芽酵母中发现的端粒位置效应(TPE)都是染色体位置对基因表达可能产生影响的例子(Gottschling et al., 1990)。基因在PEV中发生差异表达是因为染色体倒位产生了新的异染色质-常染色质边界,而与异染色质区域并置的常染色质基因经历了异染色质诱导的基因沉默(Hessler, 1958;埃尔金,路透社,2013)。染色体位置对基因表达的影响在拟南芥45S rDNA位点的表达中得到了很好的研究(Mohannath et al., 2016)。此外,其他研究表明,由于大型调控结构域的重组,基因表达的变化伴随着染色体重排的引入,例如倒位或易位(Naseeb et al., 2016)。据报道,它们也会引起断点附近遗传区域的修饰(Lavington &;Kern, 2017),易位和邻近区域的表观遗传环境(Wesley &;ean, 1994;Fournier et al., 2010),或引起核重组(Fournier et al., 2010;Harewood et al., 2010)。然而,目前尚不清楚所报道的基因表达和表观遗传变化是在引入染色体重排后立即发生的,还是随着时间的推移在后代中建立起来的。为了揭示染色体倒位对拟沙蚕染色质表观遗传状态和基因活性的影响,我们采用CRISPR/ cas辅助染色体工程技术,生成了两个不同大小的染色体倒位(Rönspies et al., 2022a)。倒置首先通过倒置连接的测序和荧光原位杂交(FISH)证实。然后,利用识别H3K4me3和H3K9me2为eu-和异色组蛋白标记的抗体,利用全基因组亚硫酸盐测序(WGBS)和染色质免疫沉淀测序(ChIP-seq),将这些品系的表观遗传状态与野生型(WT)植物进行比较。最后,分析了染色体重排对基因活性的影响。我们的研究结果表明,除了轻微的全基因组影响外,所研究的倒位染色体片段及其邻近区域都没有改变表观遗传标记和基因表达,这表明CRISPR/ cas诱导的染色体重组后的表观基因组和转录组具有稳健性,至少在后代中是这样。
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来源期刊
New Phytologist
New Phytologist 生物-植物科学
自引率
5.30%
发文量
728
期刊介绍: New Phytologist is an international electronic journal published 24 times a year. It is owned by the New Phytologist Foundation, a non-profit-making charitable organization dedicated to promoting plant science. The journal publishes excellent, novel, rigorous, and timely research and scholarship in plant science and its applications. The articles cover topics in five sections: Physiology & Development, Environment, Interaction, Evolution, and Transformative Plant Biotechnology. These sections encompass intracellular processes, global environmental change, and encourage cross-disciplinary approaches. The journal recognizes the use of techniques from molecular and cell biology, functional genomics, modeling, and system-based approaches in plant science. Abstracting and Indexing Information for New Phytologist includes Academic Search, AgBiotech News & Information, Agroforestry Abstracts, Biochemistry & Biophysics Citation Index, Botanical Pesticides, CAB Abstracts®, Environment Index, Global Health, and Plant Breeding Abstracts, and others.
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