• <tr id="yyy80"></tr>
  • <sup id="yyy80"></sup>
  • <tfoot id="yyy80"><noscript id="yyy80"></noscript></tfoot>
  • 99热精品在线国产_美女午夜性视频免费_国产精品国产高清国产av_av欧美777_自拍偷自拍亚洲精品老妇_亚洲熟女精品中文字幕_www日本黄色视频网_国产精品野战在线观看 ?

    Role of Long Non-coding RNAs in Reprogramming to Induced Pluripotency

    2020-07-29 05:34:36ShahzinaKanwalXiangpengGuoCarlWardGiacomoVolpeBaomingQinMiguelEstebanXichenBao
    Genomics,Proteomics & Bioinformatics 2020年1期

    Shahzina Kanwal ,Xiangpeng Guo ,Carl Ward ,Giacomo Volpe ,Baoming Qin ,Miguel A.Esteban ,6,*,Xichen Bao

    1Joint School of Life Sciences,Guangzhou Medical University and Guangzhou Institutes of Biomedicine and Health,Guangzhou 511436,China

    2Key Laboratory of Regenerative Biology and Guangdong Provincial Key Laboratory of Stem Cells and Regenerative Medicine,Guangzhou Institutes of Biomedicine and Health,Chinese Academy of Sciences,Guangzhou 510530,China

    3Laboratory of RNA,Chromatin,and Human Disease,Guangzhou Institutes of Biomedicine and Health,Chinese Academy of Sciences,Guangzhou 510530,China

    4Guangzhou Regenerative Medicine and Health Guangdong Laboratory(GRMH-GDL),Guangzhou 510005,China

    5Laboratory of Metabolism and Cell Fate,Guangzhou Institutes of Biomedicine and Health,Chinese Academy of Sciences,Guangzhou 510530,China

    6Institute for Stem Cells and Regeneration,Chinese Academy of Sciences,Beijing 100101,China

    7Laboratory of RNA Molecular Biology,Guangzhou Institutes of Biomedicine and Health,Chinese Academy of Sciences,Guangzhou 510530,China

    KEYWORDS Mesenchymal-to-epithelial transition;X chromosome reactivation;Apoptosis;Proliferation

    Abstract The generation of induced pluripotent stem cells through somatic cell reprogramming requires a global reorganization of cellular functions.This reorganization occurs in a multi-phased manner and involves a gradual revision of both the epigenome and transcriptome.Recent studies have shown that the large-scale transcriptional changes observed during reprogramming also apply to long noncoding RNAs(lncRNAs),a type of traditionally neglected RNA species that are increasingly viewed as critical regulators of cellular function.Deeper understanding of lncRNAs in reprogramming may not only help to improve this process but also have implications for studying cell plasticity in other contexts,such as development,aging,and cancer.In this review,we summarize the current progress made in profiling and analyzing the role of lncRNAs in various phases of somatic cell reprogramming,with emphasis on the re-establishment of the pluripotency gene network and X chromosome reactivation.

    Introduction

    High-throughput sequencing techniques have demonstrated that mammalian genomes are ubiquitously transcribed[1].The transcription of large numbers of non-coding RNAs(ncRNAs)[2]might help to explain interspecies differences despite limited variations in the number and sequence of coding genes.ncRNAs are classified based on their length into small RNAs(<200 nucleotides),e.g.,microRNAs(miRNAs)and piwi-interacting RNAs (piRNAs), and long ncRNAs(lncRNAs,>200 nucleotides)[3,4].

    lncRNAs are typically transcribed by RNA polymerase II,although this process occurs with different modalities and origins.They include circular RNAs(circRNAs)[5],enhancer RNAs(eRNAs)[6],antisense transcripts[7],and long intergenic ncRNAs(lincRNAs)[8,9].These lncRNAs recruit epigenetic regulators to chromatin and serve as scaffolds to stabilize protein complexes or as decoys for proteins and miRNAs[10-12].These diverse regulatory modes are due to the complementary base pairing of lncRNAs with both DNA and other RNAs,the ability of lncRNAs to interact with proteins,and the localization of lncRNAs in different cellular compartments(e.g.,cytoplasm,nucleus,or mitochondrion)[13,14].Interestingly,a large fraction of lncRNAs also associate with ribosomes and, paradoxically, some previously annotated lncRNAs produce small peptides with biological functions[15,16].The latter raises relevant questions regarding the true nature of some lncRNAs,but the general significance of these findings requires further investigation.

    Although the functional relevance of lncRNAs has not yet been systematically explored,some are known to regulate key aspects of normal and pathological cellular functions such as proliferation[17],metabolism[18,19],and epithelial cytoarchitecture[20].The recent discovery that lncRNAs display cellspecific and development-specific expression patterns has also suggested a pivotal role of lncRNAs in cell fate determination[21-23].In support of this idea,suppression of several embryonic stem cell(ESC)-specific lincRNAs influences pluripotency maintenance/exit and early lineage commitment[21].Similarly,the heart-associated lncRNA Braveheart serves as a key player in the activation of the core vascular gene network during mouse cardiac cell fate specification[22].In addition,lincRNA Yin Yang 1(linc-YY1),which is highly conserved in humans,regulates myogenesis through dislodgement of the YY1/polycomb repressive complex on target promoters,resulting in the activation of muscle gene expression[23].

    Here,we summarize the current knowledge of lncRNA profiling and functions in an extreme scenario of cell fate transition,somatic cell reprogramming.

    lncRNAs are new players in somatic cell reprogramming

    Mammalian somatic cells can be reprogrammed to an ESC state and this has revolutionized stem cell research[24,25].Originally, the reprogramming factor cocktail contained SOX2,KLF4,OCT4(encoded by Pou5f1 or POU5F1 gene in mice or humans,respectively),and c-MYC(SKOM),but other combinations of exogenous factors are also effective[26,27]. More recently, mouse reprogramming has been achieved using only chemicals [28]. Remarkably, induced pluripotent stem cells(iPSCs)provide a priori unlimited number of individual-specific stem cells that can be used for in vitro disease modeling, drug screening, and potential cell-based therapies[29,30].Although producing iPSCs from different cell sources and mammalian species is in general no longer an issue[31-34],the underlying mechanisms remain unclear and clarifying them is important for improving iPSC quality[35].

    Reprogramming occurs through a stepwise process that involves reorganization of most cell functions,culminating in the reactivation of the pluripotency gene program[36,37].This conversion is characterized by several roadblocks and checkpoints.For example,reprogramming cells must overcome the senescence/apoptosis barrier to acquire the ability to proliferate indefinitely[38,39],switch their metabolism from oxidative phosphorylation to glycolysis [40], and undergo a mesenchymal-to-epithelial transition(MET)[41,42].By the end of these events,under standard conditions,only a small percentage of the original population activates the endogenous pluripotency gene circuitry.

    Since the first demonstration of somatic cell reprogramming,multiple regulatory factors have been identified.Among these,miRNAs play essential roles in creating or removing reprogramming roadblocks[43,44].For example,components of miRNA cluster 302-367 suppress Tgfbr2 to neutralize the pro-mesenchymal effects of TGFβ cytokines secreted by somatic cells or present in serum, facilitating the MET[45,46].miRNA cluster 302-367 also targets genes encoding chromatin regulators(e.g.,BAF170)that prevent the activation of pluripotency genes in the late phase of reprogramming[46].Conceivably,lncRNAs,which are present in much higher numbers than miRNAs and are in principle more versatile players in cell regulation,could be important regulators of reprogramming too.In this regard,lncRNAs also experience phase-dependent changes during reprogramming,and their regulation shares the same epigenetic mechanisms that drive mRNA changes in reprogramming[47,48].These findings suggest a coordinated role between protein-coding and ncRNAs in reorganizing cellular functions.Studying lncRNAs is,therefore,important to understand reprogramming as a whole(Table 1).Such knowledge may also contribute to clarifying the role of lncRNAs in other contexts such as development,aging,and cancer.

    p53-regulated lncRNAs in reprogramming

    Proliferation facilitates the appearance of stochastic events needed for directing chromatin reorganization into a pluripotent state during reprogramming[38].However,somatic cells have a limited life span/proliferation capacity,and reprogramming is a stressful process involving activation of senescence/-cell death pathways.Accordingly,suppressing p53,p16Ink4a,or p19Arfenhances reprogramming efficiency by accelerating proliferation and/or reducing apoptosis[39,49,50].

    Interestingly,several p53-regulated lncRNAs have strong impacts on reprogramming efficiency(Figure 1).For instance,expression of lincRNA-RoR(regulator of reprogramming)[51]and lincRNA-p21[52,53]is induced by p53[54,55],which can influence reprogramming in positive and negative manner,respectively. Similarly, expression of p53-repressedLNCPRESS1(lncRNA p53-regulated and ESC-associated 1)is robustly induced during reprogramming and activates the pluripotency network[56].Given these circumstances,it would be useful to systematically profile lncRNAs during reprogramming in the presence or absence of p53 or other pro-senescence regulators.

    Table 1 lncRNAs and their functions in somatic cell reprogramming

    lincRNA-RoR

    lincRNA-RoR(2603 nucleotides)is located on chromosome 18q21.31.Its expression is induced by p53[54]and facilitates human reprogramming by suppressing the p53-mediated transcriptional response to oxidative stress and DNA damage[51].However,overexpression or knockdown of lincRNA-RoR does not influence cell growth at the early stages of reprogramming.In cancer cells, lincRNA-RoR suppresses p53 translation through heterogeneous nuclear ribonucleoprotein I(hnRNP I),a classical RNA-binding protein(RBP)involved in splicing[54].In this regard,a fraction of hnRNP I is localized in the cytoplasm,where it promotes the translation of p53 through binding to internal ribosome entry sites.Apart from suppressing the p53 pathway, lincRNA-RoR sequesters prodifferentiation miRNAs,including miR-145,to maintain the expression of the core pluripotency transcription factors OCT4,SOX2,and NANOG in human ESCs[57,58].Although these two latter mechanisms have not yet been tested in reprogramming,it is likely that they also contribute to the effects of lincRNA-RoR in this process.Notably,lincRNA-RoR is more highly expressed in human iPSCs than in ESCs[51],which may be a consequence of a selective advantage conferred during reprogramming.

    lincRNA-p21

    lincRNA-p21 (3121 nucleotides, located on chromosome 6p21.2)resides 5 kb upstream of p21.It was originally discovered as an executioner of the p53-mediated apoptotic response[55].Expression of lincRNA-p21 is induced by p53 and impairs mouse reprogramming,but different cis or trans modes of action have been proposed[52,53].The cis-acting model is based on the observation that conditional excision of the lincRNA-p21 promoter and first exon reduces the expression of p21 mRNA,resulting in enhanced proliferation and reprogramming efficiency based on alkaline phosphatase activity(an early marker of reprogramming)[53].However,it is important to note that removal of the lincRNA-p21 locus leads to the loss of multiple enhancers that control the transcription of nearby genes,including p21 itself,independently of lincRNA-p21[59].In fact,removal of the lincRNA-p21 locus alters the expression of nearby genes even in tissues with no detectable lincRNA-p21 transcript.Conversely,the trans-acting model of lincRNA-p21 in reprogramming proposes that lincRNA-p21 impairs reprogramming independently of proliferation by binding to pluripotency loci(e.g.,Nanog)and blocking their reactivation[52].In this model,lincRNA-p21 represses pluripotency loci by recruiting the histone 3 lysine 9(H3K9)methyltransferase SETDB1 or the maintenance DNA methyltransferase DNMT1.Binding of lincRNA-p21 to these epigenetic regulators is mediated by the classical RBP hnRNP-K.Hence,suppressing hnRNP-K also results in enhanced reprogramming efficiency, although it is unlikely that it acts exclusively through lincRNA-p21. The idea that expression of p53-induced lncRNAs such as lincRNA-p21 prevents reprogramming independent of proliferation or apoptosis is attractive,as this may have implications for the control of cell fate when stem cells are under stress or in aging and in p53-negative cancers.

    In a screen for lncRNAs modulating the conversion of preiPSCs to iPSCs,lincRNA-p21 was identified as a negative regulator of reprogramming[52].Pre-iPSCs are stable but incompletely reprogrammed clones,and their conversion to iPSCs is commonly used as a proxy for the late phase of reprogramming[60].The same screen also identified lincRNA-1463 and lincRNA-1526 as negative regulators of the conversion of pre-iPSCs to iPSCs.However,it is unclear whether expression of these two lncRNAs is also regulated by p53.

    LNCPRESS1

    Another screening study conducted on differentiating human ESCs led to the discovery of LNCPRESS1(832 nucleotides,located on chromosome 7q22.1). LNCPRESS1 is a p53-repressed lncRNA that positively regulates the pluripotency gene network in human ESCs[56].LNCPRESS1 acts as a decoy for the histone deacetylase SIRT6,thereby enriching the activating H3K56/K9 acetylation marks at pluripotency loci. Interestingly, expression of LNCPRESS1 is strongly induced during reprogramming[56],suggesting a potential role of LNCPRESS1 in improving reprogramming efficiency.

    Other lncRNAs regulating reprogramming

    In addition to lncRNAs regulated by p53, several other lncRNAs have been identified which can regulate somatic cell reprogramming through MET,metabolic switch,or reactivation of the pluripotency gene network(Figure 1).ESCs are epithelial-like,therefore,reprogramming of mesenchymal-like cells such as fibroblasts to iPSCs unavoidably involves the acquisition of an epithelial phenotype.This occurs in the early phase of reprogramming through MET [41,42,61]. Since pluripotent cells and somatic cells have different metabolic profiles, reprogramming, logically, also requires metabolic remodeling.iPSCs exhibit a metabolic shift from oxidative phosphorylation to glycolysis, which is associated with a decreased number and complexity of mitochondria.Importantly,blocking MET or the metabolic switch derails reprogramming [41,42,62], but not every reprogramming intermediate that successfully passes through these checkpoints achieves full activation of the pluripotency gene network. Likewise, even when somatic cells have been successfully reprogrammed to iPSCs,their DNA methylation patterns often do not faithfully reflect those of ESCs.For example, iPSCs can show aberrant DNA methylation of imprinted regions,which alters the expression of lncRNAs encoded by those regions[63-65].

    Figure 1 lncRNAs regulating somatic cell reprogramming

    Zeb2-NAT

    Zinc finger E-box binding homeobox 2-natural antisense transcript(Zeb2-NAT,430 nucleotides,located on chromosome 2q22.3)is a natural antisense transcript for the transcription factor ZEB2 [66]. ZEB2 is a master regulator of the epithelial-to-mesenchymal transition (EMT) that represses the expression of many epithelial genes[67].Zeb2-NAT is involved in maintaining Zeb2 expression by preventing the splicing of the Zeb2 5′-untranslated region(5′-UTR)[66].Interestingly,Zeb2-NAT is highly expressed in fibroblasts from aged mice[68],which are known to be less amenable to reprogramming[69].The reprogramming efficiency of these aged fibroblasts is enhanced when expression of Zeb2-NAT is reduced. Given that aging promotes the accumulation of DNA damage,thereby activating p53,it is plausible that expression of Zeb2-NAT is also induced by p53 in aged fibroblasts and during reprogramming.However,this speculation has not yet been tested.

    Gas5

    Expression of the lncRNA growth arrest specific 5(Gas5,656 nucleotides,located on chromosome 1q25.1)is controlled by pluripotency transcription factors.Gas5 plays a pivotal role in reprogramming and self-renewal of mouse ESCs by maintaining the expression of genes encoding key pluripotency factors and ten-eleven translocation 1(Tet1)[70].The effect of Gas5 on Tet1 expression suggests that Gas5 regulates active DNA demethylation in reprogramming.Its human ortholog,GAS5,also controls human ESC pluripotency by protecting NODAL mRNA from miRNA-mediated degradation[71],a mechanism that may also participate in human reprogramming.GAS5 is also a well-known regulator of cell proliferation and apoptosis in various cell contexts,including breast cancer,lung cancer,and differentiating mouse ESCs[70,72,73].

    Ladr lncRNAs

    Single-cell transcriptomic analysis of mouse reprogramming has unveiled numerous lncRNAs that are activated or repressed during this process[47].Among them,knockdown of two lncRNAs activated during reprogramming 49 and 83(Ladr49 and Ladr83),showed modest effects on reprogramming efficiency,but led to the upregulation of muscle-related genes in reprogramming intermediate cells.Interestingly,these two lncRNAs have been previously shown to physically associate with polycomb repressive complex 2(PRC2)[21,74].On the other hand,depletion of Ladr86 and Ladr91,which show upregulated expression in reprogramming, represses mitochondria-associated genes,suggesting a role in the metabolic switch during reprogramming.

    Promoter/enhancer-interacting lncRNAs

    A recently devised approach chromatin-RNA in situ reversetranscription sequencing(CRIST-seq)took advantage of the specificity of the clustered regularly interspaced short palindromic repeats(CRISPR)/CRISPR-associated 9(Cas9)system for DNA to profile pluripotency-specific lncRNAs that interact with the promoter of the core pluripotency genes Sox2 and Pou5f1[75].Among the identified lncRNAs interacting with the Sox2 promoter, 59 of them were differentially expressed in reprogramming.Notably,overexpression of one of these differentially expressed lncRNAs,Sox2 promoterinteracting lncRNA 14(Spilr14,also known as Snhg14),led to significant enhancement of reprogramming efficiency,whereas its knockdown caused loss of pluripotency in iPSCs.Another lncRNA, Pou5f1 enhancer-binding lncRNA 20(Peblr20),was identified utilizing a strategy combining RNA reverse transcription-associated capture sequencing (RATseq)and RNA sequencing[76].Peblr20 is expressed at higher levels in iPSCs than in mouse embryonic fibroblasts and promotes reprogramming by activating endogenous Pou5f1 in trans through the recruitment of TET2 to the enhancer region,which enhances the expression of eRNAs.

    Imprinted lncRNAs

    Maternally expressed 3(Meg3,also known as Gtl2)is localized at the imprinted Dlk1-Dio3 gene cluster on mouse chromosome 12qF1,which also encodes numerous miRNAs[77,78].ncRNAs harbored in this region are maternally expressed in mammals.Interestingly,they are strongly repressed in most mouse iPSCs compared to ESCs,which is responsible for the failure to support the development of all-iPSC mice[63,79].This inability can be reversed through reactivation of the imprinted Dlk1-Dio3 locus in reprogramming using the histone deacetylase inhibitor valproic acid or ascorbic acid(vitamin C, Vc) [63,80,81]. Vc facilitates the conservation of imprinting at this locus by interfering with reprogramming factor-induced loss of H3K4 methylation,which prevents the recruitment of DNA methyltransferase 3A (DNMT3A).Importantly,MEG3,which is encoded in the DLK1-DIO3 locus,is also frequently silenced in human iPSCs[82].Similarly,hypomethylation of the maternally expressed imprinted lncRNA zinc finger(CCCH type),RNA binding motif,and serine/arginine rich 1 (Zrsr1) is associated with reduced pluripotency in mouse iPSCs,which could not be rescued by treating reprogramming cells with Vc or PD0325901 and CHIR99021(inhibitors of MEK1/2 and GSK3,respectively)[64].It remains to be clarified how these imprinted lncRNAs regulate the reprogramming process.

    X chromosome reactivation during reprogramming

    In the late stage of reprogramming,another important process involving lncRNAs is X chromosome reactivation (XCR)(Figure 2). During the development of mice and other mammals, one of the two X chromosomes in females is randomly silenced in somatic cells shortly after implantation to maintain the dosage equivalence between the sexes[83,84].X chromosome inactivation(XCI)is initiated by the expression of X-inactive specific transcript(Xist),a 17,918-nucleotide lncRNA localized in the X chromosome inactivation center(XIC)[85].Xist coats the X chromosome,which gradually removes RNA polymerase II and active histone marks such as H3K4 trimethylation.This is followed by a sequential gain of diverse repressive marks including H3K27 trimethylation (H3K27me3), macroH2A.1 histone(macroH2A),and DNA methylation on the inactivated X chromosome(Xi).Consequently,Xi is silenced throughout life with remarkable stability.Despite possessing a stable nature,Xi can be reactivated in specific contexts such as primordial germ cell differentiation,somatic cell nuclear transfer(SCNT),and somatic cell reprogramming[86].Due to its ease and reproducibility, reprogramming provides an unprecedented tool for characterizing the events involved in XCR[87,88].

    A relevant question in the field of reprogramming is the order of the epigenetic events leading to XCR. Highresolution single-cell time course analyses of reprogramming using immunofluorescence and RNA fluorescence in situ hybridization(FISH)[87,89]demonstrate that XCR occurs in the following order:(1)recruitment of H3K27 methyltransferase EZH2 to Xi(XiEZH2+)in E-cadherin(CDH1)positive(CDH1+)cells,(2)activation of NANOG in XiEZH2+cells,(3)loss of EZH2,H3K27me3,Xist coating,and macroH2A1 on Xi in NANOG+cells,and(4)removal of DNA methylation on Xi and activation of the transcribed antisense to Xist(Tsix).Therefore,except for DNA demethylation,the events of XCR in reprogramming follow the inverse order of the events of XCI in development. Notably, the removal of DNA methylation during XCR in reprogramming is TETindependent and hence passive,and the expression of Tsix is paradoxically dispensable.

    Conspicuously,despite the initiation of XCR in NANOG+cells,full reactivation of the core pluripotency circuitry precedes XCR in reprogramming.Accordingly,XCR serves as one of the crucial standards for bona fide mouse iPSC identification[90].This is consistent with the observation that the pluripotency factor PRDM14 represses Xist expression[91].Although ectopic expression of Xist impairs XCR in reprogramming,its depletion does not affect XCR or reprogramming efficiency [87]. This is inconsistent with the observations in SCNT, the efficiency of which is greatly improved following Xist elimination[92].A possible explanation could be that Xist plays opposite roles at different stages of somatic cell reprogramming,as SCNT has no obvious phases.In support of this idea,Xist depletion impairs the MET in the early phase of reprogramming but significantly improves the conversion of pre-iPSCs to iPSCs[93].Interestingly,the reprogramming booster Vc[81]promotes XCR in reprogramming by preventing the relocalization of macroH2A onto Xi[93].This is likely mediated by the boosting effect of Vc on H3K27me3 demethylases[94,95].It remains to be determined whether Vc also acts through unrelated mechanisms,for example by decreasing the levels of N6-methyladenosine(m6A)deposition on Xist,as Vc is also a cofactor for another two dioxygenases,the alpha-ketoglutarate-dependent dioxygenase fat mass and obesity-associated protein (FTO) and AlkB homolog 5(ALKBH5),which are responsible for erasing this epitranscriptomic mark[96-98].

    Notably,the aforementioned mechanisms have been investigated in mouse cell reprogramming,but it is unclear whether the same principles apply to human cells.In this regard,XCR is either absent or unstable in female donors when human iPSCs are generated using standard culture conditions[99],but is present in reprogramming to na?¨ve pluripotency[100].Further studies are needed to understand the differences and similarities between XCR in mouse and human cell reprogramming.

    Perspectives

    Figure 2 X chromosome reactivation in somatic cell reprogramming

    It is becoming increasingly evident that lncRNAs are critical players in cell fate regulation.Few lncRNAs,mostly related to p53 or cell senescence,have been well studied thus far,but the repertoire of lncRNAs that regulate pluripotency/reprogramming is likely large.Systematic profiling is needed for functional characterization of these lncRNAs.This could be facilitated by developing highly efficient/deterministic reprogramming systems[101],as low-efficiency protocols are prone to cell heterogeneity and the cell reprogramming kinetics are asynchronous,which complicates the interpretations.In addition, specialized high-throughput RNA sequencing approaches such as global nuclear run-on sequencing(GROseq)and sequencing with RNase R treatment are necessary to enrich for certain lncRNAs such as eRNAs and circRNAs[102,103],respectively,since they are rarely detected using conventional RNA sequencing methodologies.In this regard,several circRNAs exhibit human iPSC/ESC-specific expression[104,105].Among them,circBIRC6 and circCORO1C positively regulate pluripotency maintenance and reprogramming[104]. circBIRC6 acts as a sponge for differentiationmediated miRNAs,whereas the mechanism underlying circCORO1C regulation is unknown.An important consideration is that functional lncRNA screens with short hairpin RNAs(shRNAs)or small interfering RNAs(siRNAs)have inevitable limitations because many lncRNAs are localized in the nucleus,where shRNAs/siRNAs cannot effectively silence transcripts.As a solution,CRISPR/Cas9-based screening systems assisted by comprehensive computer prediction algorithms could be used to increase on-target efficiency and minimize off-target effects.This approach requires eliminating the whole DNA fragment(rather than introducing a frame shift for coding genes),potentially in the promoter region,to suppress expression/functionality of lncRNAs without affecting the nearby genes[106].CRISPR/Cas13 and CRISPR interference are potential alternatives,as they can be used to edit RNA or repress gene expression without altering the genome[107,108].Collectively,the characterization of lncRNAs in reprogramming will not only help to uncover new layers of regulation for the diverse pathways modulating reprogramming but may also have implications in other types of cell fate transitions.

    Competing interests

    The authors have declared no competing interests.

    Acknowledgments

    We thank all members of the Esteban lab for their support.Research in the authors’laboratories is supported by the National Key R&D Program of China (Grant Nos.2016YFA0100701,2016YFA0100102,2018YFA0106903,and 2016YFA0100300);the Strategic Priority Research Program of the Chinese Academy of Sciences (Grant No.XDA16030502);the Youth Innovation Promotion Association of the Chinese Academy of Sciences(Grant No.2015294),the National Natural Science Foundation of China(Grant Nos.31671537, 31571524, and 31501192); the Natural Science Foundation of Guangdong Province (Grant No.2018B030306042),the Guangdong Province Science and Technology Program (Grant Nos. 2014A030312001,2016A050503037, 2016B030229007, and 2017B050506007),the Science and Technology Planning Project of Guangdong Province (Grant No. 2017B030314056), the Pearl River Science and Technology Nova Program of Guangzhou(Grant No.201610010107),and the Guangzhou Science and Technology Program(Grant No.201807010066),China.SK is supported by a President’s International Fellowship Initiative program from the Chinese Academy of Sciences and CW is supported by a Pearl River Overseas Young Talents Postdoctoral Fellowship.

    ORCID

    0000-0003-4694-136X(Kanwal S)

    0000-0002-3323-1134(Guo X)

    0000-0003-0889-9025(Ward C)

    0000-0001-5000-6951(Volpe G)

    0000-0001-7652-161X(Qin B)

    0000-0002-1426-6809(Esteban MA)

    0000-0003-0389-4233(Bao X)

    国产淫语在线视频| 午夜日本视频在线| 国产黄色视频一区二区在线观看| 美女中出高潮动态图| 男人和女人高潮做爰伦理| 一区二区三区四区激情视频| 久久97久久精品| 午夜免费鲁丝| 久久青草综合色| 嫩草影院入口| av在线播放精品| 久久午夜福利片| 亚洲精品久久午夜乱码| 亚洲欧美日韩卡通动漫| 大片免费播放器 马上看| 毛片一级片免费看久久久久| 六月丁香七月| 亚洲精品国产色婷婷电影| 少妇人妻久久综合中文| av福利片在线| 日韩中字成人| 在线看a的网站| 国产精品福利在线免费观看| 免费av不卡在线播放| 亚洲国产精品一区二区三区在线| 亚洲久久久国产精品| 欧美高清成人免费视频www| 观看免费一级毛片| 人妻人人澡人人爽人人| 国产综合精华液| 日本av手机在线免费观看| 男女国产视频网站| 婷婷色综合大香蕉| 精品一区二区免费观看| 亚洲自偷自拍三级| 嫩草影院新地址| 大香蕉97超碰在线| 51国产日韩欧美| 热99国产精品久久久久久7| 狠狠精品人妻久久久久久综合| 男人狂女人下面高潮的视频| 2022亚洲国产成人精品| 日本-黄色视频高清免费观看| 国产熟女欧美一区二区| 成人毛片a级毛片在线播放| 久久国产精品大桥未久av | 亚洲,一卡二卡三卡| 国产av国产精品国产| av一本久久久久| 国产精品女同一区二区软件| 日日摸夜夜添夜夜爱| 水蜜桃什么品种好| av黄色大香蕉| 日韩一区二区视频免费看| 免费人成在线观看视频色| 少妇高潮的动态图| 中文字幕人妻丝袜制服| 伊人亚洲综合成人网| 日本午夜av视频| 精品久久久久久电影网| 我要看日韩黄色一级片| 国模一区二区三区四区视频| 美女视频免费永久观看网站| a 毛片基地| av免费在线看不卡| 午夜免费鲁丝| 狂野欧美激情性bbbbbb| 嫩草影院新地址| 亚洲国产色片| 色视频www国产| 特大巨黑吊av在线直播| 国产在线一区二区三区精| 久久亚洲国产成人精品v| 亚洲国产精品一区二区三区在线| 深夜a级毛片| 免费不卡的大黄色大毛片视频在线观看| 青青草视频在线视频观看| 我要看黄色一级片免费的| 国产乱来视频区| 在线观看人妻少妇| 内射极品少妇av片p| 亚洲欧美一区二区三区黑人 | 国产成人91sexporn| av一本久久久久| 亚洲,一卡二卡三卡| 黄色视频在线播放观看不卡| 亚洲精品国产av蜜桃| 日韩电影二区| 亚洲中文av在线| 在线看a的网站| 99热国产这里只有精品6| 一级片'在线观看视频| 一级毛片久久久久久久久女| 一区二区三区乱码不卡18| 秋霞在线观看毛片| 99久久精品国产国产毛片| 在线亚洲精品国产二区图片欧美 | 国产乱人偷精品视频| 亚洲国产精品国产精品| 十八禁高潮呻吟视频 | 国产精品久久久久久久电影| 亚洲第一区二区三区不卡| 日韩一本色道免费dvd| 日韩av在线免费看完整版不卡| 国产一区二区在线观看日韩| 日韩中文字幕视频在线看片| 天堂8中文在线网| 一级片'在线观看视频| 观看美女的网站| 久久青草综合色| 日本av手机在线免费观看| 99久久综合免费| 热99国产精品久久久久久7| 最近2019中文字幕mv第一页| 最近中文字幕高清免费大全6| 久久久国产一区二区| 国产精品免费大片| 国产亚洲欧美精品永久| 搡老乐熟女国产| 中文字幕精品免费在线观看视频 | 亚洲精品久久久久久婷婷小说| 日韩一本色道免费dvd| 丝袜喷水一区| 国产日韩欧美亚洲二区| 午夜精品国产一区二区电影| 少妇 在线观看| 国模一区二区三区四区视频| 女性被躁到高潮视频| 日韩 亚洲 欧美在线| 在线免费观看不下载黄p国产| 热99国产精品久久久久久7| 久久人人爽人人片av| 精品人妻熟女av久视频| 欧美3d第一页| 在线观看www视频免费| 国产精品一二三区在线看| 国产视频内射| 亚洲精品456在线播放app| 久久99热6这里只有精品| 久久人人爽人人片av| 亚洲图色成人| 成年女人在线观看亚洲视频| 久久久久久久国产电影| 99热这里只有是精品50| www.av在线官网国产| 国产精品一区二区性色av| 男女啪啪激烈高潮av片| 国产熟女午夜一区二区三区 | 成人黄色视频免费在线看| 免费av中文字幕在线| 亚洲怡红院男人天堂| 最新的欧美精品一区二区| 国产av一区二区精品久久| 午夜视频国产福利| 国产毛片在线视频| 国产男女超爽视频在线观看| 国产白丝娇喘喷水9色精品| 成人亚洲精品一区在线观看| 亚洲欧美精品自产自拍| freevideosex欧美| 亚洲av国产av综合av卡| 一区二区av电影网| 男女边吃奶边做爰视频| 少妇猛男粗大的猛烈进出视频| 麻豆成人午夜福利视频| 欧美丝袜亚洲另类| 精品酒店卫生间| 一级毛片 在线播放| 大香蕉97超碰在线| kizo精华| 久久国产乱子免费精品| 嫩草影院入口| 国产色婷婷99| 亚洲精品久久久久久婷婷小说| 久久久久久久精品精品| 久久久国产欧美日韩av| 中文字幕人妻丝袜制服| 欧美另类一区| 国产精品久久久久久精品古装| 欧美最新免费一区二区三区| 免费久久久久久久精品成人欧美视频 | 国产日韩欧美亚洲二区| 精品人妻偷拍中文字幕| 搡老乐熟女国产| 特大巨黑吊av在线直播| 亚洲电影在线观看av| 久久久国产一区二区| 久久精品国产亚洲网站| 日韩,欧美,国产一区二区三区| 一区二区av电影网| 人妻少妇偷人精品九色| 高清视频免费观看一区二区| 久久久久久伊人网av| 曰老女人黄片| 国产成人一区二区在线| 日韩制服骚丝袜av| 男人爽女人下面视频在线观看| 秋霞伦理黄片| 亚洲精品久久久久久婷婷小说| 亚洲色图综合在线观看| 丝袜喷水一区| 亚洲美女黄色视频免费看| 欧美日韩国产mv在线观看视频| 美女中出高潮动态图| 国产真实伦视频高清在线观看| 美女内射精品一级片tv| 人妻 亚洲 视频| av免费观看日本| 久久精品久久精品一区二区三区| 精品少妇久久久久久888优播| 久久久精品免费免费高清| 久久国产精品男人的天堂亚洲 | 亚洲自偷自拍三级| 一区二区三区免费毛片| 在线观看www视频免费| 亚洲国产欧美日韩在线播放 | 99久久精品热视频| 亚洲情色 制服丝袜| 亚洲欧美一区二区三区国产| 久久久久久久久久久丰满| 欧美+日韩+精品| 一区在线观看完整版| 少妇裸体淫交视频免费看高清| 最近的中文字幕免费完整| 少妇的逼水好多| 91午夜精品亚洲一区二区三区| 少妇的逼好多水| 亚洲婷婷狠狠爱综合网| 老司机亚洲免费影院| 麻豆精品久久久久久蜜桃| 久久久a久久爽久久v久久| 亚洲国产日韩一区二区| 成人毛片60女人毛片免费| 国产成人91sexporn| 少妇裸体淫交视频免费看高清| 一本一本综合久久| 精品久久久噜噜| 免费观看av网站的网址| 大码成人一级视频| 日韩亚洲欧美综合| 亚洲三级黄色毛片| 天天操日日干夜夜撸| 一级片'在线观看视频| 曰老女人黄片| 成人午夜精彩视频在线观看| 久久热精品热| 成人影院久久| 热re99久久精品国产66热6| 日韩,欧美,国产一区二区三区| 日本午夜av视频| 大片免费播放器 马上看| 亚洲国产精品国产精品| 精品熟女少妇av免费看| 五月玫瑰六月丁香| 亚洲精品国产av蜜桃| a级毛片在线看网站| 卡戴珊不雅视频在线播放| 国产欧美日韩精品一区二区| 婷婷色综合大香蕉| 久久精品夜色国产| 精品熟女少妇av免费看| 久久国产亚洲av麻豆专区| 在线观看一区二区三区激情| 最新中文字幕久久久久| 在线免费观看不下载黄p国产| 大陆偷拍与自拍| 亚洲国产精品一区三区| 美女中出高潮动态图| 国内揄拍国产精品人妻在线| 人妻系列 视频| 欧美三级亚洲精品| 男男h啪啪无遮挡| 久久久欧美国产精品| 十分钟在线观看高清视频www | 波野结衣二区三区在线| 午夜免费鲁丝| 日韩成人伦理影院| 久久久精品94久久精品| 成人综合一区亚洲| 色婷婷av一区二区三区视频| 特大巨黑吊av在线直播| 街头女战士在线观看网站| 国产老妇伦熟女老妇高清| 女人久久www免费人成看片| 青春草亚洲视频在线观看| 特大巨黑吊av在线直播| 免费高清在线观看视频在线观看| 精品一区二区三区视频在线| 免费观看在线日韩| 国模一区二区三区四区视频| 成人免费观看视频高清| 一区二区三区四区激情视频| 伦精品一区二区三区| 国产亚洲午夜精品一区二区久久| 日本欧美视频一区| 七月丁香在线播放| 99热这里只有精品一区| 国产午夜精品久久久久久一区二区三区| 国内精品宾馆在线| 欧美日韩综合久久久久久| 亚洲情色 制服丝袜| 美女中出高潮动态图| 色吧在线观看| 乱人伦中国视频| 五月伊人婷婷丁香| 韩国高清视频一区二区三区| 九九在线视频观看精品| 亚洲欧美日韩卡通动漫| 伊人久久精品亚洲午夜| 观看美女的网站| 亚洲精品一二三| 男女边吃奶边做爰视频| 在线观看一区二区三区激情| 爱豆传媒免费全集在线观看| 观看免费一级毛片| 国产精品无大码| 丰满迷人的少妇在线观看| 久久婷婷青草| 久久久国产一区二区| 久久久a久久爽久久v久久| 国产熟女午夜一区二区三区 | 久久久久久久久久人人人人人人| 久久久精品94久久精品| 日韩免费高清中文字幕av| 国国产精品蜜臀av免费| 亚洲国产成人一精品久久久| 国产真实伦视频高清在线观看| 亚洲精品第二区| 中文天堂在线官网| 在线 av 中文字幕| 亚洲不卡免费看| 精品久久久久久久久亚洲| 久久99热6这里只有精品| 91午夜精品亚洲一区二区三区| 在线 av 中文字幕| 大话2 男鬼变身卡| 在线 av 中文字幕| 纵有疾风起免费观看全集完整版| 男人和女人高潮做爰伦理| 国产熟女欧美一区二区| 毛片一级片免费看久久久久| 黄色一级大片看看| 一级二级三级毛片免费看| 另类亚洲欧美激情| av天堂久久9| h视频一区二区三区| 中文欧美无线码| 建设人人有责人人尽责人人享有的| 国产永久视频网站| 国产精品国产三级国产av玫瑰| 午夜精品国产一区二区电影| 69精品国产乱码久久久| 亚洲精品国产av蜜桃| 日韩中字成人| 精品一区二区三区视频在线| 国产免费一级a男人的天堂| 中文字幕人妻丝袜制服| 黑人猛操日本美女一级片| 91久久精品国产一区二区成人| 国产探花极品一区二区| 一边亲一边摸免费视频| 青春草视频在线免费观看| 汤姆久久久久久久影院中文字幕| 99久久综合免费| 久久女婷五月综合色啪小说| 亚洲精品色激情综合| 亚洲国产毛片av蜜桃av| 日本与韩国留学比较| av在线播放精品| 亚洲国产日韩一区二区| 丰满乱子伦码专区| kizo精华| 欧美日韩视频高清一区二区三区二| 一个人免费看片子| 少妇被粗大猛烈的视频| 亚洲av在线观看美女高潮| 精品人妻偷拍中文字幕| 国产乱来视频区| 蜜桃在线观看..| 精品久久久久久久久av| 精品一品国产午夜福利视频| 精品久久久久久电影网| 欧美3d第一页| 69精品国产乱码久久久| av国产久精品久网站免费入址| 亚洲国产毛片av蜜桃av| 少妇的逼好多水| 成年人午夜在线观看视频| 亚洲av欧美aⅴ国产| 最近最新中文字幕免费大全7| 免费黄频网站在线观看国产| 亚洲精品自拍成人| 青青草视频在线视频观看| 少妇人妻精品综合一区二区| 日韩欧美一区视频在线观看 | 欧美日韩一区二区视频在线观看视频在线| 亚洲中文av在线| 波野结衣二区三区在线| 国国产精品蜜臀av免费| 精品人妻熟女av久视频| 国产精品一区二区三区四区免费观看| 亚洲熟女精品中文字幕| 久久国产亚洲av麻豆专区| 日韩欧美一区视频在线观看 | av播播在线观看一区| 3wmmmm亚洲av在线观看| 亚洲情色 制服丝袜| 免费黄色在线免费观看| 夜夜爽夜夜爽视频| 青春草亚洲视频在线观看| 亚洲国产欧美日韩在线播放 | 国产精品嫩草影院av在线观看| 国内精品宾馆在线| 蜜桃久久精品国产亚洲av| a级毛片在线看网站| 能在线免费看毛片的网站| 亚洲在久久综合| 精品国产乱码久久久久久小说| 欧美成人午夜免费资源| 国产黄频视频在线观看| 晚上一个人看的免费电影| 永久免费av网站大全| 国产精品一区www在线观看| 九草在线视频观看| 一本久久精品| 久热这里只有精品99| 七月丁香在线播放| 久久婷婷青草| 大码成人一级视频| 欧美高清成人免费视频www| 99久久精品热视频| 精品一区二区三卡| 中国国产av一级| 欧美精品高潮呻吟av久久| 国产精品三级大全| 纯流量卡能插随身wifi吗| 亚洲av成人精品一二三区| 国产中年淑女户外野战色| 国产女主播在线喷水免费视频网站| 97精品久久久久久久久久精品| 成年美女黄网站色视频大全免费 | 在线免费观看不下载黄p国产| 午夜免费观看性视频| 成人影院久久| 黄色一级大片看看| 综合色丁香网| 美女脱内裤让男人舔精品视频| 成年人午夜在线观看视频| 老司机影院毛片| 大香蕉97超碰在线| 永久网站在线| 亚洲四区av| 伊人亚洲综合成人网| 自拍欧美九色日韩亚洲蝌蚪91 | 熟女电影av网| 亚洲精品久久午夜乱码| 老司机影院毛片| 丰满乱子伦码专区| 国产在线免费精品| 亚洲av电影在线观看一区二区三区| 新久久久久国产一级毛片| 在线观看三级黄色| 国产高清三级在线| 亚洲精品,欧美精品| 成年av动漫网址| 久久国产乱子免费精品| 91成人精品电影| 成人国产麻豆网| 成人美女网站在线观看视频| 人妻人人澡人人爽人人| 麻豆成人av视频| 91久久精品国产一区二区三区| 亚洲性久久影院| 各种免费的搞黄视频| 久久毛片免费看一区二区三区| 久久久久久久久久久久大奶| 在线观看免费高清a一片| 制服丝袜香蕉在线| 最近中文字幕高清免费大全6| 两个人免费观看高清视频 | 我要看日韩黄色一级片| 在线观看免费视频网站a站| 综合色丁香网| av.在线天堂| 国产亚洲91精品色在线| a级一级毛片免费在线观看| 嫩草影院新地址| 国产精品久久久久久久电影| 人妻人人澡人人爽人人| 汤姆久久久久久久影院中文字幕| 久久精品国产亚洲av天美| 深夜a级毛片| 中文字幕人妻熟人妻熟丝袜美| 只有这里有精品99| 国语对白做爰xxxⅹ性视频网站| 久久久亚洲精品成人影院| 在线观看免费日韩欧美大片 | 一区二区三区乱码不卡18| 天天躁夜夜躁狠狠久久av| 国产亚洲一区二区精品| 全区人妻精品视频| 国产精品一二三区在线看| 亚洲av成人精品一二三区| 丝袜在线中文字幕| 欧美精品国产亚洲| 日本黄色日本黄色录像| 免费久久久久久久精品成人欧美视频 | 国产一区二区在线观看日韩| 亚洲国产毛片av蜜桃av| 七月丁香在线播放| 久久亚洲国产成人精品v| videossex国产| 高清视频免费观看一区二区| 久久 成人 亚洲| 边亲边吃奶的免费视频| 美女内射精品一级片tv| 亚洲精品视频女| 中文字幕制服av| 国产一区二区在线观看日韩| 精品视频人人做人人爽| 免费观看性生交大片5| 国产成人一区二区在线| 夜夜骑夜夜射夜夜干| 91午夜精品亚洲一区二区三区| 熟女人妻精品中文字幕| 少妇人妻一区二区三区视频| 国产无遮挡羞羞视频在线观看| 这个男人来自地球电影免费观看 | 日产精品乱码卡一卡2卡三| 亚洲国产成人一精品久久久| 两个人的视频大全免费| 色视频www国产| 成人国产av品久久久| 如日韩欧美国产精品一区二区三区 | 菩萨蛮人人尽说江南好唐韦庄| 亚洲丝袜综合中文字幕| 毛片一级片免费看久久久久| 欧美人与善性xxx| 国内精品宾馆在线| 哪个播放器可以免费观看大片| 国产精品99久久99久久久不卡 | 看免费成人av毛片| 国产亚洲精品久久久com| 国产黄频视频在线观看| 亚洲精品中文字幕在线视频 | 校园人妻丝袜中文字幕| 伦理电影大哥的女人| 亚洲高清免费不卡视频| 全区人妻精品视频| 国产高清有码在线观看视频| 亚洲无线观看免费| 国产一区亚洲一区在线观看| 天堂俺去俺来也www色官网| av网站免费在线观看视频| 国产精品久久久久成人av| 两个人的视频大全免费| 曰老女人黄片| 久久久午夜欧美精品| 欧美日韩av久久| 日产精品乱码卡一卡2卡三| 性色av一级| 91久久精品电影网| a 毛片基地| 久久久久国产网址| av国产精品久久久久影院| 精品亚洲成a人片在线观看| 欧美精品人与动牲交sv欧美| 黄色怎么调成土黄色| 岛国毛片在线播放| 最黄视频免费看| 国产精品嫩草影院av在线观看| 久久精品熟女亚洲av麻豆精品| 美女主播在线视频| 欧美日韩av久久| 国产精品偷伦视频观看了| 午夜91福利影院| 国产免费福利视频在线观看| 国产日韩一区二区三区精品不卡 | 国产欧美日韩综合在线一区二区 | 美女xxoo啪啪120秒动态图| 日韩电影二区| 好男人视频免费观看在线| 久久久久国产网址| 日韩 亚洲 欧美在线| 国产免费一级a男人的天堂| 九九爱精品视频在线观看| 一级a做视频免费观看| av在线老鸭窝| 高清不卡的av网站| 一级,二级,三级黄色视频| 高清在线视频一区二区三区| 大码成人一级视频| 国产在线视频一区二区| 亚洲欧美中文字幕日韩二区| 精品国产一区二区久久| 国产 精品1| 欧美bdsm另类| 欧美 日韩 精品 国产| 久久久久久久久大av| 不卡视频在线观看欧美| 欧美日韩视频高清一区二区三区二| 极品教师在线视频| √禁漫天堂资源中文www| 久久久精品免费免费高清| 伦理电影免费视频| 欧美日韩一区二区视频在线观看视频在线| 亚洲av电影在线观看一区二区三区| 男女免费视频国产| 久久久精品94久久精品| 国产精品一区二区三区四区免费观看| 麻豆乱淫一区二区| 人妻夜夜爽99麻豆av| 国产 精品1| 91aial.com中文字幕在线观看| 秋霞伦理黄片|