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Live cell imaging of genomic loci using dCas9-SunTag system and a bright fluorescent protein

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Protein & Cell 2017年第11期8卷 853-855页

作者： Huiying Ye zhili rong Ying Lin Cancer Research Institute School of Basic Medical SciencesSouthern Medical University Guangzhou 510515 China

Dear Editor,CRISPR-Cas9 （clustered regularly interspaced short palin- dromic repeats-CRISPR associated） systems have been harnessed for kinds of genome manipulation, including gene editing, transcription regulation, and chromosome loci imaging （Dominguez et al., 2016; Komor et al., 2017）. A typical engineered CRISPR-Cas9 system is composed of a Cas9 protein and a single guide RNA （sgRNA）, which could form a protein/RNA complex to recognize and cleave DNA sequence （Hsu et al., 2014; Wright et al., 2016）.

关键词： Live cell imaging genomic loci using dCas9-SunTag system a bright fluorescent protein

Homologous recombination in human embryonic stem cells using CRISPR/Cas9 nickase and a long DNA donor template

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Protein & Cell 2014年第4期5卷 258-260页

作者： zhili rong Shengyun Zhu Yang Xu Xuemei Fu Division of Biological Sciences University of CaliforniaSan Diego9500 Gilman DriveLa JollaCA 92093USA Shenzhen Children’s Hospital 7019 Yitian RoadShenzhen 518026China Chongqing Medical University Chongqing 400016China

Dear Editor Genome editing of human embryonic stem cells （hESCs） is critical for basic biological research and regenerative medi- cine. However, until a few years ago, gene targeting tech- nologies to disrupt, repair or overexpress genes in hESCs had been very inefficient and thus could not be routinely used. Recent technical breakthrough includes bacterial arti- ficial chromosome based high efficiency gene targeting （Song et al., 2010）, as well as the successful engineering of two systems of site-specific nucleases, zinc-finger nucleases （ZFNs） and transcription activator-like effector nucleases （TALENs）（Hockemeyer et al., 2009; Hockemeyer et al.,

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Genetic approach to track neural cell fate decisions using human embryonic stem cells

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Protein & Cell 2014年第1期5卷 69-79页

作者： Xuemei Fu zhili rong Shengyun Zhu Xiaocheng Wang Yang Xu Blue B. Lake Shenzhen Children's Hospital Shenzhen 518026 China Section of Molecular Biology Division of Biological Sciences University of California San Diego 9500 Gilman Drive LaJolla CA 92093-0322 USA

With their capability to undergo unlimited self-renewal and to differentiate into all cell types in the body, human embryonic stem cells （hESCs） hold great promise in human cell therapy. However, there are limited tools for easily identifying and isolating live hESC-derived cells. To track hESC-derived neural progenitor cells （NPCs）, we applied homologous recombination to knock-in the mCherry gene into the Nestin locus of hESCs. This facilitated the genetic labeling of Nestin positive neural progenitor cells with mCherry. Our reporter system enables the visualization of neural induction from hESCs both in vitro （embryoid bodies） and in vivo （ter- atomas）. This system also permits the identification of different neural subpopulations based on the intensity of our fluorescent reporter. In this context, a high level of mCherry expression showed enrichment for neural progenitors, while lower mCherry corresponded with more committed neural states. Combination of mCherry high expression with cell surface antigen staining enabled further enrichment of hESC-derived NPCs. These mCherry＊ NPCs could be expanded in culture and their differentiation resulted in a down-regulation of mCherry consistent with the loss of Nestin expression. Therefore, we have developed a fluorescent reporter system that can be used to trace neural differentiation events of hESCs.

关键词： Nestin knock-in human embryonic stemcells neural progenitor cells

Lung adeno-squamous carcinoma modeling using oncogenic KRAS activation in human embryonic stem cell-derived alveolar organoids

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Genes & Diseases 2023年第1期10卷 33-36页

作者： Jianqi Feng Lian Li Zhihong Tan Shanshan Zhao Xin Zhang Jinhong Wang Yaping Ye Ying Lin zhili rong Cancer Research Institute School of Basic Medical SciencesState Key Laboratory of Organ Failure ResearchNational Clinical Research Center of Kidney DiseaseKey Laboratory of Organ Failure Research(Ministry of Education)Southern Medical UniversityGuangzhouGuangdong 510515China Dermatology Hospital Southern Medical UniversityGuangzhouGuangdong 510091China Department of Respiration The Third Affiliated Hospital of Southern Medical UniversityGuangzhouGuangdong 510630China Department of Pathology Nanfang Hospital and School of Basic Medical ScienceSouthern Medical UniversityGuangzhouGuangdong 510515China Bioland Laboratory(Guangzhou Regenerative Medicine and Health Guangdong Laboratory) GuangzhouGuangdong 510005China Experimental Education/Administration Center School of Basic Medical ScienceSouthern Medical UniversityGuangzhouGuangdong 510515China

Mutant KRAs is a common driver in lung ***,the occurrence of human lung cancer and its subsequent development after activation of oncogenic KRAS in epithelial cells remain poorly *** are embedded in three-dimensional Matrigel in which tissue-derived human adult stem cells can efficiently grow for a long time,maintaining genetic and phenotypic stability."But until now,the use of organoids to simulate human non-small cell lung cancer has not been reported.

关键词： lung KRAS Lung

Correction to: Gene activation in human cells using CRISPR/Cpf1-p300 and CRISPR/Cpf1- SunTag systems

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Protein & Cell 2019年第10期10卷 776-777页

作者： Xin Zhang Wei Wang Lin Shan Le Han Shufeng Ma Yan Zhang Bingtao Hao Ying Lin zhili rong Cancer Research Institute School of Basic Medical SciencesSouthern Medical UniversityGuangzhou 510515China

In the original publication the Supplementary Material and Fig.2 are in *** correct versi on of Supplementary Material and Fig.2 are provided in this correction *** text HBG2 appearing in the article should be read as... 详细信息

关键词： CRISPR/Cpf1 activation correction

Gene activation in human cells using CRISPR/ Cpf1-p300 and CRISPR/Cpf1-SunTag systems

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Protein & Cell 2018年第4期9卷 380-383页

Dear Editor, Clustered regularly interspaced short palindromic repeats （CRISPR） system is part of the adaptive immunity of bacteria and archaea that defends them against phage infection （Barrangou and Doudna, 2016）. Variant CRISPR systems have been identified and harnessed for a wide range of applications in various organisms, exemplified by gene editing with the CRISPR/Cas9 system from Streptococcuspyogenes （Barrangou and Doudna, 2016）.

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OSMRβmutants enhance basal keratinocyte differentiation via inactivation of the STAT5/KLF7 axis in PLCA patients

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Protein & Cell 2021年第8期12卷 653-660页

作者： Jun Liu Junchen ChenYadan Zhong Xiaoling Yu Ping Lu Jianqi Feng Xin Zhang Shufeng Ma Chao Yang Bin Yang zhili rong Dermatology Hospital Southern Medical UniversityGuangzhou 510091China Cancer Research Institute School of Basic Medical SciencesSouthern Medical UniversityGuangzhou 510515China Bioland Laboratory(Guangzhou Regenerative Medicine and Health Guangdong Laboratory) Guangzhou 510005China

Dear Editor,Primary localized cutaneous amyloidosis(PLCA)is a skinlimited disorder characterized by deposition of amyloid material in the superficial *** to clinical characteristics,PLCA is divided into lichen,macular,and nodular *** is found worldwide but has a higher incidence in South America and Southeast Asia,such as in Brazil and China(Chang et al.,2014;Tey et al.,2016).

关键词： al. superficial amyloid

Cell-cell contact-induced gene editing/activation in mammalian cells using a synNotch-CRISPR/Cas9 system

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Protein & Cell 2020年第4期11卷 299-303页

作者： Hongxin Huang Xin Zhang Jie Lv Hongcheng Yang Xinlong Wang Shufeng Ma Ruoyang Shao Xin Peng Ying Lin zhili rong School of Basic Medical Sciences Cancer Research InstituteSouthern Medical UniversityGuangzhou 510515China Dermatology Hospital Southern Medical UniversityGuangzhou 510091China Guangzhou Regenerative Medicine and Health Guangdong Laboratory Guangzhou 510005China

Dear Editor,The CRISPR system has been widely used for genome manipulation in various cells,tissues and whole *** an increasing variety of inducible CRISPR systems have been exploited for a variety of applications,such as chemical switch(Zetsche et al.,2015),photo switch(Shao et al.,2018)and solution ligand switch(Baeumler et al.,2017;Kipniss et al.,2017;Schwarz et al.,2017)systems,a cell-cell interaction inducible system is *** synthetic Notch(synNotch)receptor is a recently developed cell-cell contact sensing platform,which contains a customized extracellular sensor module,a transmembrane core domain of native Notch,and a customized intracellular responder module(Morsut et al.,2016).

关键词： CRISPR/Cas9 system custom

Host metabolism dysregulation and cell tropism identification in human airway and alveolar organoids upon SARS-CoV-2 infection

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Protein & Cell 2021年第9期12卷 717-733页

作者： rongjuan Pei Jianqi Feng Yecheng Zhang Hao Sun Lian Li Xuejie Yang Jiangping He Shuqi Xiao Jin Xiong Ying Lin Kun Wen Hongwei Zhou Jiekai Chen zhili rong Xinwen Chen Cancer Research Institute School of Basic Medical SciencesSouthern Medical UniversityGuangzhou 510515China Center for Biosafety Mega-Science Wuhan Institute of VirologyChinese Academy of SciencesWuhan 430071China Bioland Laboratory(Guangzhou Regenerative Medicine and Health Guangdong Laboratory) Guangzhou 510005China Dermatology Hospital Southern Medical UniversityGuangzhou 510091China Guangzhou Institutes of Biomedicine and Health Chinese Academy of SciencesGuangzhou 510530China The Centre of Cell Lineage and Atlas(CCLA) Bioland Laboratory(Guangzhou Regenerative Medicine and Health-Guangdong Laboratory)Guangzhou 510530China Joint School of Life Sciences Guangzhou Medical University and Guangzhou Institutes of Biomedicine and HealthChinese Academy of SciencesGuangzhou 511436China Microbiome Medicine Center Division of Laboratory MedicineZhujiang HospitalSouthern Medical UniversityGuangzhouChina

The coronavirus disease 2019(COVID-19)pandemic is caused by infection with the severe acute respiratory syndrome coronavirus 2(SARS-CoV-2),which is spread primary via respiratory droplets and infects the *** widely used cell lines and animals are unable to accurately mimic human physiological conditions because of the abnormal status of cell lines(transformed or cancer cells)and species differences between animals and *** are stem cell-derived selforganized three-dimensional culture in vitro and model the physiological conditions of natural *** we showed that SARS-CoV-2 infected and extensively replicated in human embryonic stem cells(hESCs)-derived lung organoids,including airway and alveolar organoids which covered the complete infection and spread route for SARS-CoV-2 within *** infected ceils were ciliated,club,and alveolar type 2(AT2)cells,which were sequentially located from the proximal to the distal airway and terminal alveoli,***,RNA-seq revealed early cell response to virus infection including an unexpected downregulation of the metabolic processes,especially lipid metabolism,in addition to the well-known upregulation of immune ***,Remdesivir and a human neutralizing antibody potently inhibited SARS-CoV-2 replication in lung ***,human lung organoids can serve as a pathophysiological model to investigate the underlying mechanism of SARS-CoV-2 infection and to discover and test therapeutic drugs for COVID-19.

关键词： COVID-19 SARS-CoV-2 lung organoids cell tropism cellular metabolism drug discovery