Blog Archive

Αλέξανδρος Γ. Σφακιανάκης

Friday, April 19, 2019

Genetics and Genomics

Perspectives on gene expression regulation techniques in Drosophila

Publication date: Available online 17 April 2019

Source: Journal of Genetics and Genomics

Author(s): Rong-Gang Xu, Xia Wang, Da Shen, Jin Sun, Huan-Huan Qiao, Fang Wang, Lu-Ping Liu, Jian-Quan Ni

Abstract

Gene expression regulation, including loss-of-function and gain-of-function assays, is a powerful method to study developmental and disease mechanisms. Drosophila melanogaster is an ideal model system particularly well-equipped with many genetic tools. In this review, we describe and discuss the gene expression regulation techniques recently developed and their applications, including the CRISPR/Cas9-triggered heritable mutation system, CRISPR/dCas9-based transcriptional activation (CRISPRa) system, and CRISPR/dCas9-based transcriptional repression (CRISPRi) system, as well as the next-generation transgenic RNAi system. The main purpose of this review is to provide the fly research community with an updated summary of newly developed gene expression regulation techniques and help the community to select appropriate methods and optimize the research strategy.



An effective strategy to establish a male sterility mutant mini-library by CRISPR/Cas9-mediated knockout of anther-specific genes in rice

Publication date: Available online 9 April 2019

Source: Journal of Genetics and Genomics

Author(s): Kun Ma, Jingluan Han, Yu Hao, Zhongfang Yang, Junyu Chen, Yao-Guang Liu, Qinlong Zhu, Letian Chen



Plant genome editing using xCas9 with expanded PAM compatibility

Publication date: Available online 5 April 2019

Source: Journal of Genetics and Genomics

Author(s): Jingying Li, Jinman Luo, Meilian Xu, Shaoya Li, Jiahui Zhang, Huiyuan Li, Lei Yan, Yunde Zhao, Lanqin Xia



Lipid storage regulator CdsA is essential for Drosophila metamorphosis

Publication date: Available online 3 April 2019

Source: Journal of Genetics and Genomics

Author(s): Yuan Liu, Yuan Ji, Xia Li, Guanghou Shui, Xun Huang



IBM1-dependent H3K9 demethylation enables self-silencing of an exogenous silencer for the non-cell autonomous silencing of an endogenous target gene

Publication date: Available online 23 March 2019

Source: Journal of Genetics and Genomics

Author(s): Xiaolan Liu, Jianhua Zhao, Hui-Shan Guo



Zfhx3 is essential for progesterone/progesterone receptor signaling to drive ductal side-branching and alveologenesis in mouse mammary glands

Publication date: Available online 22 March 2019

Source: Journal of Genetics and Genomics

Author(s): Gui Ma, Ang Gao, Yinan Yang, Yuan He, Xi Zhang, Baotong Zhang, Zhiqian Zhang, Mei Li, Xing Fu, Dan Zhao, Rui Wu, Leilei Qi, Qingxia Hu, Juan Li, Liya Fu, Zhengmao Zhu, Jin-Tang Dong

Abstract

Progesterone (Pg)/progesterone receptor (PR) signaling drives mammary gland side-branching and alveologenesis, but the mechanisms through which Pg/PR signaling functions remain to be clarified. Using in vitro and in vivo models and histological and molecular analyses, we determined the role of Zfhx3 transcription factor in mammary gland development driven by Pg/PR signaling. Postnatal deletion of Zfhx3 in mouse mammary epithelial cells attenuated side-branching morphogenesis and alveologenesis. These effects were undetectable in the absence of Pg/PR signaling. During the estrus cycle, Zfhx3 expression corresponded to that of Pg, being at the highest level at the diestrus stage; Zfhx3 deletion inhibited mammary gland branching more potently at diestrus than estrus stage. Loss of Zfhx3 not only attenuated the expansion of stem/progenitor cells driven by Pg/PR signaling, but also impaired the function of Pg/PR signaling in the transcriptional activation of multiple genes. In addition, Pg/PR signaling significantly expanded PR- and Zfhx3-positive epithelial cells, and induced the physical association of ZFHX3 with PR. These findings establish Zfhx3 as an integral transcription factor of Pg/PR signaling in driving side-branching and alveologenesis during mammary gland development.



Arabidopsis AGC protein kinases IREH1 and IRE3 control root skewing

Publication date: Available online 22 March 2019

Source: Journal of Genetics and Genomics

Author(s): Xiaozhen Yue, Zhiai Guo, Teng Shi, Lizhen Song, Youfa Cheng

Abstract

AGC protein kinases play important roles in plant growth and development. Several AGC kinases in Arabidopsis have been functionally characterized. However, the "AGC Other" subfamily, including IRE, IREH1, IRE3 and IRE4, has not been well understood. Here, we reported that ireh1 mutants displayed a root skewing phenotype, which can be enhanced by ire3 mutation. IREH1 and IRE3 were expressed in roots, consistent with their function in controlling root skewing. The fluorescence intensities of the microtubule marker KNpro:EGFP-MBD were decreased in ireh1ire3 and ireh1 ire3 mutants compared to wild type. The microtubule arrangements in ireh1 and ireh1 ire3 mutants were also altered. IREH1 physically interacted with IRE3 in vitro and in planta. Thus, our findings demonstrate that IREH1 and IRE3 protein kinases play important roles in controlling root skewing, and maintaining microtubule network in Arabidopsis.



CPI-17-mediated contraction of vascular smooth muscle is essential for the development of hypertension in obese mice

Publication date: Available online 15 March 2019

Source: Journal of Genetics and Genomics

Author(s): Jie Sun, Tao Tao, Wei Zhao, Lisha Wei, Fan She, Pei Wang, Yeqiong Li, Yanyan Zheng, Xin Chen, Wei Wang, Yanning Qiao, Xue-Na Zhang, Min-Sheng Zhu

Abstract

Several factors have been implicated in obesity-related hypertension, but the genesis of the hypertension is largely unknown. In this study, we found a significantly upregulated expression of CPI-17 (C-kinase-potentiated protein phosphatase 1 inhibitor of 17 kDa) and protein kinase C (PKC) isoforms in the vascular smooth muscles of high-fat diet (HFD)-fed obese mice. The obese wild-type mice showed a significant elevation of blood pressure and enhanced calcium-sensitized contraction of vascular smooth muscles. However, the obese CPI-17-deficient mice showed a normotensive blood pressure, and the calcium-sensitized contraction was consistently reduced. In addition, the mutant muscle displayed an abolished responsive force to a PKC activator and a 30%–50% reduction in both the initial peak force and sustained force in response to various G protein-coupled receptor (GPCR) agonists. Our observations showed that CPI-17-mediated calcium sensitization is mediated through a GPCR/PKC/CPI-17/MLCP/RLC signaling pathway. We therefore propose that the upregulation of CPI-17-mediated calcium-sensitized vasocontraction by obesity contributes to the development of obesity-related hypertension.



PIK-75 promotes homology-directed DNA repair

Publication date: Available online 15 March 2019

Source: Journal of Genetics and Genomics

Author(s): Guoling Li, Xianwei Zhang, Hao Ou, Haoqiang Wang, Dewu Liu, Huaqiang Yang, Zhenfang Wu



SAA-Cas9: A tunable genome editing system with increased bio-safety and reduced off-target effects

Publication date: Available online 15 March 2019

Source: Journal of Genetics and Genomics

Author(s): Sanyuan Ma, Jianduo Zhang, Wei Lu, Yue Liu, Qingyou Xia



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