An inducible CRISPR-ON system for controllable gene activation in human pluripotent stem cells.


:Human pluripotent stem cells (hPSCs) are an important system to study early human development, model human diseases, and develop cell replacement therapies. However, genetic manipulation of hPSCs is challenging and a method to simultaneously activate multiple genomic sites in a controllable manner is sorely needed. Here, we constructed a CRISPR-ON system to efficiently upregulate endogenous genes in hPSCs. A doxycycline (Dox) inducible dCas9-VP64-p65-Rta (dCas9-VPR) transcription activator and a reverse Tet transactivator (rtTA) expression cassette were knocked into the two alleles of the AAVS1 locus to generate an iVPR hESC line. We showed that the dCas9-VPR level could be precisely and reversibly controlled by the addition and withdrawal of Dox. Upon transfection of multiplexed gRNA plasmid targeting the NANOG promoter and Dox induction, we were able to control NANOG gene expression from its endogenous locus. Interestingly, an elevated NANOG level promoted naïve pluripotent gene expression, enhanced cell survival and clonogenicity, and enabled hESCs to integrate with the inner cell mass (ICM) of mouse blastocysts in vitro. Thus, iVPR cells provide a convenient platform for gene function studies as well as high-throughput screens in hPSCs.


Protein Cell


Protein & cell


Guo J,Ma D,Huang R,Ming J,Ye M,Kee K,Xie Z,Na J




Has Abstract


2017-05-01 00:00:00














  • Oncogenic miR-19a and miR-19b co-regulate tumor suppressor MTUS1 to promote cell proliferation and migration in lung cancer.

    abstract::MTUS1 (microtubule-associated tumor suppressor 1) has been identified that can function as a tumor suppressor gene in many malignant tumors. However, the function and mechanisms underlying the regulation of MTUS1 are unclear. In the present study, we reported that miR-19a and miR-19b (miR-19a/b) promote proliferation ...

    journal_title:Protein & cell

    pub_type: 杂志文章


    authors: Gu Y,Liu S,Zhang X,Chen G,Liang H,Yu M,Liao Z,Zhou Y,Zhang CY,Wang T,Wang C,Zhang J,Chen X

    更新日期:2017-06-01 00:00:00

  • An examination of the OMIM database for associating mutation to a consensus reference sequence.

    abstract::Gene mutation (e.g. substitution, insertion and deletion) and related phenotype information are important biomedical knowledge. Many biomedical databases (e.g. OMIM) incorporate such data. However, few studies have examined the quality of this data. In the current study, we examined the quality of protein single-point...

    journal_title:Protein & cell

    pub_type: 杂志文章


    authors: Li Z,Ying B,Liu X,Zhang X,Yu H

    更新日期:2012-03-01 00:00:00

  • Dephosphorylation of cGAS by PPP6C impairs its substrate binding activity and innate antiviral response.

    abstract::The cyclic GMP-AMP (cGAMP) synthase (cGAS) plays a critical role in host defense by sensing cytosolic DNA derived from microbial pathogens or mis-located cellular DNA. Upon DNA binding, cGAS utilizes GTP and ATP as substrates to synthesize cGAMP, leading to MITA-mediated innate immune response. In this study, we ident...

    journal_title:Protein & cell

    pub_type: 杂志文章


    authors: Li M,Shu HB

    更新日期:2020-08-01 00:00:00

  • Complement activation by phospholipids: the interplay of factor H and C1q.

    abstract::Complement proteins in blood recognize charged particles. The anionic phospholipid (aPL) cardiolipin binds both complement proteins C1q and factor H. C1q is an activator of the complement classical pathway, while factor H is an inhibitor of the alternative pathway. To examine opposing effects of C1q and factor H on co...

    journal_title:Protein & cell

    pub_type: 杂志文章


    authors: Tan LA,Yu B,Sim FC,Kishore U,Sim RB

    更新日期:2010-11-01 00:00:00

  • Switch of substrate specificity of hyperthermophilic acylaminoacyl peptidase by combination of protein and solvent engineering.

    abstract::The inherent evolvability of promiscuous enzymes endows them with great potential to be artificially evolved for novel functions. Previously, we succeeded in transforming a promiscuous acylaminoacyl peptidase (apAAP) from the hyperthermophilic archaeon Aeropyrum pernix K1 into a specific carboxylesterase by making a s...

    journal_title:Protein & cell

    pub_type: 杂志文章


    authors: Liu C,Yang G,Wu L,Tian G,Zhang Z,Feng Y

    更新日期:2011-06-01 00:00:00

  • The role of Smad7 in oral mucositis.

    abstract::Oral mucositis, a severe oral ulceration, is a common toxic effect of radio- or chemoradio-therapy and a limiting factor to using the maximum dose of radiation for effective cancer treatment. Among cancer patients, at least 40% and up to 70%, of individuals treated with standard chemotherapy regimens or upper-body rad...

    journal_title:Protein & cell

    pub_type: 杂志文章,评审


    authors: Bian L,Han G,Zhao CW,Garl PJ,Wang XJ

    更新日期:2015-03-01 00:00:00

  • Arrestins: structural disorder creates rich functionality.

    abstract::Arrestins are soluble relatively small 44-46 kDa proteins that specifically bind hundreds of active phosphorylated GPCRs and dozens of non-receptor partners. There are binding partners that demonstrate preference for each of the known arrestin conformations: free, receptor-bound, and microtubule-bound. Recent evidence...

    journal_title:Protein & cell

    pub_type: 杂志文章,评审


    authors: Gurevich VV,Gurevich EV,Uversky VN

    更新日期:2018-12-01 00:00:00

  • Thymic epithelial cell development and differentiation: cellular and molecular regulation.

    abstract::Thymic epithelial cells (TECs) are one of the most important components in thymic microenvironment supporting thymocyte development and maturation. TECs, composed of cortical and medullary TECs, are derived from a common bipotent progenitor, mediating thymocyte positive and negative selections. Multiple levels of sign...

    journal_title:Protein & cell

    pub_type: 杂志文章,评审


    authors: Sun L,Luo H,Li H,Zhao Y

    更新日期:2013-05-01 00:00:00

  • UV-B-induced photomorphogenesis in Arabidopsis.

    abstract::Ultraviolet-B (UV-B) is a relatively minor component of sunlight, but can induce stress-related physiological processes or UV-B-specific photomorphogenic responses in plants. In the last decade, significant progress has been made in understanding the UV-B photomorphogenic pathway, including identification of the key c...

    journal_title:Protein & cell

    pub_type: 杂志文章,评审


    authors: Li J,Yang L,Jin D,Nezames CD,Terzaghi W,Deng XW

    更新日期:2013-07-01 00:00:00

  • Genome engineering of stem cell organoids for disease modeling.

    abstract::Precision medicine emerges as a new approach that takes into account individual variability. Successful realization of precision medicine requires disease models that are able to incorporate personalized disease information and recapitulate disease development processes at the molecular, cellular and organ levels. Wit...

    journal_title:Protein & cell

    pub_type: 杂志文章,评审


    authors: Sun Y,Ding Q

    更新日期:2017-05-01 00:00:00

  • Cells derived from iPSC can be immunogenic - yes or no?

    abstract::The induced pluripotent stem cells (iPSCs), derived by ectopic expression of reprogramming factors in somatic cells, can potentially provide unlimited autologous cells for regenerative medicine. In theory, the autologous cells derived from patient iPSCs should be immune tolerant by the host without any immune rejectio...

    journal_title:Protein & cell

    pub_type: 杂志文章


    authors: Cao J,Li X,Lu X,Zhang C,Yu H,Zhao T

    更新日期:2014-01-01 00:00:00

  • Core pluripotency factors promote glycolysis of human embryonic stem cells by activating GLUT1 enhancer.

    abstract::Human embryonic stem cells (hESCs) depend on glycolysis for energy and substrates for biosynthesis. To understand the mechanisms governing the metabolism of hESCs, we investigated the transcriptional regulation of glucose transporter 1 (GLUT1, SLC2A1), a key glycolytic gene to maintain pluripotency. By combining the g...

    journal_title:Protein & cell

    pub_type: 杂志文章


    authors: Yu L,Ji KY,Zhang J,Xu Y,Ying Y,Mai T,Xu S,Zhang QB,Yao KT,Xu Y

    更新日期:2019-09-01 00:00:00

  • Domesticated cynomolgus monkey embryonic stem cells allow the generation of neonatal interspecies chimeric pigs.

    abstract::Blastocyst complementation by pluripotent stem cell (PSC) injection is believed to be the most promising method to generate xenogeneic organs. However, ethical issues prevent the study of human chimeras in the late embryonic stage of development. Primate embryonic stem cells (ESCs), which have similar pluripotency to ...

    journal_title:Protein & cell

    pub_type: 杂志文章


    authors: Fu R,Yu D,Ren J,Li C,Wang J,Feng G,Wang X,Wan H,Li T,Wang L,Zhang Y,Hai T,Li W,Zhou Q

    更新日期:2020-02-01 00:00:00

  • Integrative analysis of in vivo recording with single-cell RNA-seq data reveals molecular properties of light-sensitive neurons in mouse V1.

    abstract::Vision formation is classically based on projections from retinal ganglion cells (RGC) to the lateral geniculate nucleus (LGN) and the primary visual cortex (V1). Neurons in the mouse V1 are tuned to light stimuli. Although the cellular information of the retina and the LGN has been widely studied, the transcriptome p...

    journal_title:Protein & cell

    pub_type: 杂志文章


    authors: Liu J,Wang M,Sun L,Pan NC,Zhang C,Zhang J,Zuo Z,He S,Wu Q,Wang X

    更新日期:2020-06-01 00:00:00

  • The Fanconi anemia pathway and DNA interstrand cross-link repair.

    abstract::Fanconi anemia (FA) is an autosomal or X-linked recessive disorder characterized by chromosomal instability, bone marrow failure, cancer susceptibility, and a profound sensitivity to agents that produce DNA interstrand cross-link (ICL). To date, 15 genes have been identified that, when mutated, result in FA or an FA-l...

    journal_title:Protein & cell

    pub_type: 杂志文章,评审


    authors: Su X,Huang J

    更新日期:2011-09-01 00:00:00

  • Phage display screening of therapeutic peptide for cancer targeting and therapy.

    abstract::Recently, phage display technology has been announced as the recipient of Nobel Prize in Chemistry 2018. Phage display technique allows high affinity target-binding peptides to be selected from a complex mixture pool of billions of displayed peptides on phage in a combinatorial library and could be further enriched th...

    journal_title:Protein & cell

    pub_type: 杂志文章,评审


    authors: Saw PE,Song EW

    更新日期:2019-11-01 00:00:00

  • New discovery rarely runs smooth: an update on progranulin/TNFR interactions.

    abstract::Progranulin (PGRN) is a growth factor implicated in various pathophysiological processes, including wound healing, inflammation, tumorigenesis, and neurodegeneration. It was previously reported that PGRN binds to tumor necrosis factor receptors (TNFR) and has therapeutic effects in inflammatory arthritis (Tang et. al,...

    journal_title:Protein & cell

    pub_type: 杂志文章,评审


    authors: Wang BC,Liu H,Talwar A,Jian J

    更新日期:2015-11-01 00:00:00

  • Crystal structures of the two membrane-proximal Ig-like domains (D3D4) of LILRB1/B2: alternative models for their involvement in peptide-HLA binding.

    abstract::Leukocyte immunoglobulin-like receptors (LILRs), also called CD85s, ILTs, or LIRs, are important mediators of immune activation and tolerance that contain tandem immunoglobulin (Ig)-like folds. There are 11 (in addition to two pseudogenes) LILRs in total, two with two Ig-like domains (D1D2) and the remaining nine with...

    journal_title:Protein & cell

    pub_type: 杂志文章


    authors: Nam G,Shi Y,Ryu M,Wang Q,Song H,Liu J,Yan J,Qi J,Gao GF

    更新日期:2013-10-01 00:00:00

  • Secreted miR-34a in astrocytic shedding vesicles enhanced the vulnerability of dopaminergic neurons to neurotoxins by targeting Bcl-2.

    abstract::MicroRNAs (miRNAs) are a class of noncoding RNAs that regulates target gene expression at posttranscriptional level, leading to further biological functions. We have demonstrated that microvesicles (MVs) can deliver miRNAs into target cells as a novel way of intercellular communication. It is reported that in central ...

    journal_title:Protein & cell

    pub_type: 杂志文章


    authors: Mao S,Sun Q,Xiao H,Zhang C,Li L

    更新日期:2015-07-01 00:00:00

  • MiR-29b suppresses the proliferation and migration of osteosarcoma cells by targeting CDK6.

    abstract::Osteosarcoma is the most common primary sarcoma of bone, and it is a leading cause of cancer death among adolescents and young adults. However, the molecular mechanism underlying osteosarcoma carcinogenesis remains poorly understood. Recently, cyclin-dependent kinase 6 (CDK6) was identified as an important oncogene. W...

    journal_title:Protein & cell

    pub_type: 杂志文章


    authors: Zhu K,Liu L,Zhang J,Wang Y,Liang H,Fan G,Jiang Z,Zhang CY,Chen X,Zhou G

    更新日期:2016-06-01 00:00:00

  • Correction to: A binding-block ion selective mechanism revealed by a Na/K selective channel.

    abstract::In the original publication the PDB numbers were not cited. ...

    journal_title:Protein & cell

    pub_type: 已发布勘误


    authors: Yu J,Zhang B,Zhang Y,Xu CQ,Zhuo W,Ge J,Li J,Gao N,Li Y,Yang M

    更新日期:2019-07-01 00:00:00

  • Structures of the N- and C-terminal domains of MHV-A59 nucleocapsid protein corroborate a conserved RNA-protein binding mechanism in coronavirus.

    abstract::Coronaviruses are the causative agent of respiratory and enteric diseases in animals and humans. One example is SARS, which caused a worldwide health threat in 2003. In coronaviruses, the structural protein N (nucleocapsid protein) associates with the viral RNA to form the filamentous nucleocapsid and plays a crucial ...

    journal_title:Protein & cell

    pub_type: 杂志文章


    authors: Ma Y,Tong X,Xu X,Li X,Lou Z,Rao Z

    更新日期:2010-07-01 00:00:00

  • Dscam mutation leads to hydrocephalus and decreased motor function.

    abstract::The nervous system is one of the most complicated organ systems in invertebrates and vertebrates. Down syndrome cell adhesion molecule (DSCAM) of the immunoglobulin (Ig) superfamily is expressed widely in the nervous system during embryonic development. Previous studies in Drosophila suggest that Dscam plays important...

    journal_title:Protein & cell

    pub_type: 杂志文章


    authors: Xu Y,Ye H,Shen Y,Xu Q,Zhu L,Liu J,Wu JY

    更新日期:2011-08-01 00:00:00

  • Transmembrane domain dependent inhibitory function of FcγRIIB.

    abstract::FcγRIIB, the only inhibitory IgG Fc receptor, functions to suppress the hyper-activation of immune cells. Numerous studies have illustrated its inhibitory function through the ITIM motif in the cytoplasmic tail of FcγRIIB. However, later studies revealed that in addition to the ITIM, the transmembrane (TM) domain of F...

    journal_title:Protein & cell

    pub_type: 杂志文章,评审


    authors: Wang J,Li Z,Xu L,Yang H,Liu W

    更新日期:2018-12-01 00:00:00

  • MicroRNAs: a new ray of hope for diabetes mellitus.

    abstract::Diabetes mellitus has become one of the most common chronic diseases, thereby posing a major challenge to global health. Characterized by high levels of blood glucose (hyperglycemia), diabetes usually results from a loss of insulin-producing β-cells in the pancreas, leading to a deficiency of insulin (type 1 diabetes)...

    journal_title:Protein & cell

    pub_type: 杂志文章,评审


    authors: Kumar M,Nath S,Prasad HK,Sharma GD,Li Y

    更新日期:2012-10-01 00:00:00

  • The genetic and epigenetic alterations in human hepatocellular carcinoma: a recent update.

    abstract::Hepatocellular carcinoma (HCC) is one of the most frequent human malignancies worldwide with very poor prognosis. It is generally accepted that the progression of HCC is a long-term process with accumulation of multiple genetic and epigenetic alterations, which further lead to the activation of critical oncogenes or i...

    journal_title:Protein & cell

    pub_type: 杂志文章,评审


    authors: Liu M,Jiang L,Guan XY

    更新日期:2014-09-01 00:00:00

  • Directed hepatic differentiation from embryonic stem cells.

    abstract::The liver is the largest internal organ in mammals, and is important for the maintenance of normal physiological functions of other tissues and organs. Hepatitis, cirrhosis, liver cancer and other chronic liver diseases are serious threats to human health, and these problems are compounded by a scarcity of liver donor...

    journal_title:Protein & cell

    pub_type: 杂志文章,评审


    authors: Chen X,Zeng F

    更新日期:2011-03-01 00:00:00

  • Ligand binding and conformational changes of SUR1 subunit in pancreatic ATP-sensitive potassium channels.

    abstract::ATP-sensitive potassium channels (KATP) are energy sensors on the plasma membrane. By sensing the intracellular ADP/ATP ratio of β-cells, pancreatic KATP channels control insulin release and regulate metabolism at the whole body level. They are implicated in many metabolic disorders and diseases and are therefore impo...

    journal_title:Protein & cell

    pub_type: 杂志文章


    authors: Wu JX,Ding D,Wang M,Kang Y,Zeng X,Chen L

    更新日期:2018-06-01 00:00:00

  • Binding of Shewanella FadR to the fabA fatty acid biosynthetic gene: implications for contraction of the fad regulon.

    abstract::The Escherichia coli fadR protein product, a paradigm/prototypical FadR regulator, positively regulates fabA and fabB, the two critical genes for unsaturated fatty acid (UFA) biosynthesis. However the scenario in the other Ɣ-proteobacteria, such as Shewanella with the marine origin, is unusual in that Rodionov and cow...

    journal_title:Protein & cell

    pub_type: 杂志文章


    authors: Zhang H,Zheng B,Gao R,Feng Y

    更新日期:2015-09-01 00:00:00

  • POST1/C12ORF49 regulates the SREBP pathway by promoting site-1 protease maturation.

    abstract::Sterol-regulatory element binding proteins (SREBPs) are the key transcriptional regulators of lipid metabolism. The activation of SREBP requires translocation of the SREBP precursor from the endoplasmic reticulum to the Golgi, where it is sequentially cleaved by site-1 protease (S1P) and site-2 protease and releases a...

    journal_title:Protein & cell

    pub_type: 杂志文章


    authors: Xiao J,Xiong Y,Yang LT,Wang JQ,Zhou ZM,Dong LW,Shi XJ,Zhao X,Luo J,Song BL

    更新日期:2020-07-14 00:00:00