Cell Regeneration最新文献

{"title":"Characterization of a novel cell penetrating peptide derived from human Oct4","authors":"Eva Harreither ,&nbsp;Hanna A Rydberg ,&nbsp;Helene L Åmand ,&nbsp;Vaibhav Jadhav ,&nbsp;Lukas Fliedl ,&nbsp;Christina Benda ,&nbsp;Miguel A Esteban ,&nbsp;Duanqing Pei ,&nbsp;Nicole Borth ,&nbsp;Regina Grillari-Voglauer ,&nbsp;Oliver Hommerding ,&nbsp;Frank Edenhofer ,&nbsp;Bengt Nordén ,&nbsp;Johannes Grillari","doi":"10.1186/2045-9769-3-2","DOIUrl":"10.1186/2045-9769-3-2","url":null,"abstract":"<div><h3>Background</h3><p>Oct4 is a transcription factor that plays a major role for the preservation of the pluripotent state in embryonic stem cells as well as for efficient reprogramming of somatic cells to induced pluripotent stem cells (iPSC) or other progenitors. Protein-based reprogramming methods mainly rely on the addition of a fused cell penetrating peptide. This study describes that Oct4 inherently carries a protein transduction domain, which can translocate into human and mouse cells.</p></div><div><h3>Results</h3><p>A 16 amino acid peptide representing the third helix of the human Oct4 homeodomain, referred to as Oct4 protein transduction domain (Oct4-PTD), can internalize in mammalian cells upon conjugation to a fluorescence moiety thereby acting as a cell penetrating peptide (CPP). The cellular distribution of Oct4-PTD shows diffuse cytosolic and nuclear staining, whereas penetratin is strictly localized to a punctuate pattern in the cytoplasm. By using a Cre/loxP-based reporter system, we show that this peptide also drives translocation of a functionally active Oct4-PTD-Cre-fusion protein. We further provide evidence for translocation of full length Oct4 into human and mouse cell lines without the addition of any kind of cationic fusion tag. Finally, physico-chemical properties of the novel CPP are characterized, showing that in contrast to penetratin a helical structure of Oct4-PTD is only observed if the FITC label is present on the N-terminus of the peptide.</p></div><div><h3>Conclusions</h3><p>Oct4 is a key transcription factor in stem cell research and cellular reprogramming. Since it has been shown that recombinant Oct4 fused to a cationic fusion tag can drive generation of iPSCs, our finding might contribute to further development of protein-based methods to generate iPSCs.</p><p>Moreover, our data support the idea that transcription factors might be part of an alternative paracrine signalling pathway, where the proteins are transferred to neighbouring cells thereby actively changing the behaviour of the recipient cell.</p></div>","PeriodicalId":9811,"journal":{"name":"Cell Regeneration","volume":"3 1","pages":"Article 3:2"},"PeriodicalIF":0.0,"publicationDate":"2014-01-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://sci-hub-pdf.com/10.1186/2045-9769-3-2","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"32824084","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Generation of multi-gene knockout rabbits using the Cas9/gRNA system","authors":"Quanmei Yan ,&nbsp;Quanjun Zhang ,&nbsp;Huaqiang Yang ,&nbsp;Qingjian Zou ,&nbsp;Chengcheng Tang ,&nbsp;Nana Fan ,&nbsp;Liangxue Lai","doi":"10.1186/2045-9769-3-12","DOIUrl":"10.1186/2045-9769-3-12","url":null,"abstract":"<div><p>The prokaryotic clustered regularly interspaced short palindromic repeat (CRISPR)-associated system (Cas) is a simple, robust and efficient technique for gene targeting in model organisms such as zebrafish, mice and rats. In this report, we applied CRISPR technology to rabbits by microinjection of Cas9 mRNA and guided RNA (gRNA) into the cytoplasm of pronuclear-stage embryos. We achieved biallelic gene knockout (KO) rabbits by injection of 1 gene (IL2rg) or 2 gene (IL2rg and RAG1) Cas9 mRNA and gRNA with an efficiency of 100%. We also tested the efficiency of multiple gene KOs in early rabbit embryos and found that the efficiency of simultaneous gene mutation on target sites is as high as 100% for 3 genes (IL2rg, RAG1 and RAG2) and 33.3% for 5 genes (IL2rg, RAG1, RAG2, TIKI1 and ALB). Our results demonstrate that the Cas9/gRNA system is a highly efficient and fast tool not only for single-gene editing but also for multi-gene editing in rabbits.</p></div>","PeriodicalId":9811,"journal":{"name":"Cell Regeneration","volume":"3 1","pages":"Article 3:12"},"PeriodicalIF":0.0,"publicationDate":"2014-01-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://sci-hub-pdf.com/10.1186/2045-9769-3-12","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"32824482","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Genetic reporter analysis reveals an expandable reservoir of OCT4+ cells in adult skin","authors":"Anne Limbourg ,&nbsp;Sabine Schnabel ,&nbsp;Vladimir J Lozanovski ,&nbsp;L Christian Napp ,&nbsp;Teng-Cheong Ha ,&nbsp;Tobias Maetzig ,&nbsp;Johann Bauersachs ,&nbsp;Hassan Y Naim ,&nbsp;Axel Schambach ,&nbsp;Florian P Limbourg","doi":"10.1186/2045-9769-3-9","DOIUrl":"10.1186/2045-9769-3-9","url":null,"abstract":"<div><p>The transcription factor <em>Oct4</em> (<em>Pou5f1</em>) is a critical regulator of pluripotency in embryonic and induced pluripotent stem cells. Therefore, <em>Oct4</em> expression might identify somatic stem cell populations with inherent multipotent potential or a propensity for facilitated reprogramming. However, analysis of <em>Oct4</em> expression is confounded by <em>Oct4</em> pseudogenes or non-pluripotency-related isoforms. Systematic analysis of a transgenic <em>Oct4-EGFP</em> reporter mouse identified testis and skin as two principle sources of <em>Oct4</em>⁺ cells in postnatal mice. While the prevalence of GFP⁺ cells in testis rapidly declined with age, the skin-resident GFP⁺ population expanded in a cyclical fashion. These cells were identified as epidermal stem cells dwelling in the stem cell niche of the hair follicle, which endogenously expressed all principle reprogramming factors at low levels. Interestingly, skin wounding or non-traumatic hair removal robustly expanded the GFP⁺ epidermal cell pool not only locally, but also in uninjured skin areas, demonstrating the existence of a systemic response. Thus, the epithelial stem cell niche of the hair follicle harbors an expandable pool of Oct4+ stem cells, which might be useful for therapeutic cell transfer or facilitated reprogramming.</p></div>","PeriodicalId":9811,"journal":{"name":"Cell Regeneration","volume":"3 1","pages":"Article 3:9"},"PeriodicalIF":0.0,"publicationDate":"2014-01-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://sci-hub-pdf.com/10.1186/2045-9769-3-9","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"32824480","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"A mitochondrial strategy for safeguarding the reprogrammed genome","authors":"Alessandro Prigione ,&nbsp;James Adjaye","doi":"10.1186/2045-9769-3-5","DOIUrl":"10.1186/2045-9769-3-5","url":null,"abstract":"<div><p>Genomic aberrations induced by somatic cell reprogramming are a major drawback for future applications of this technology in regenerative medicine. A new study by Ji et al. published in Stem Cell Reports suggests a counteracting strategy based on balancing the mitochondrial/oxidative stress pathway through antioxidant supplementation.</p></div>","PeriodicalId":9811,"journal":{"name":"Cell Regeneration","volume":"3 1","pages":"Article 3:5"},"PeriodicalIF":0.0,"publicationDate":"2014-01-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://sci-hub-pdf.com/10.1186/2045-9769-3-5","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"32824476","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Aberrant DNA methylation reprogramming during induced pluripotent stem cell generation is dependent on the choice of reprogramming factors","authors":"Aline C Planello ,&nbsp;Junfeng Ji ,&nbsp;Vivek Sharma ,&nbsp;Rajat Singhania ,&nbsp;Faridah Mbabaali ,&nbsp;Fabian Müller ,&nbsp;Javier A Alfaro ,&nbsp;Christoph Bock ,&nbsp;Daniel D De Carvalho ,&nbsp;Nizar N Batada","doi":"10.1186/2045-9769-3-4","DOIUrl":"10.1186/2045-9769-3-4","url":null,"abstract":"<div><p>The conversion of somatic cells into pluripotent stem cells via overexpression of reprogramming factors involves epigenetic remodeling. DNA methylation at a significant proportion of CpG sites in induced pluripotent stem cells (iPSCs) differs from that of embryonic stem cells (ESCs). Whether different sets of reprogramming factors influence the type and extent of aberrant DNA methylation in iPSCs differently remains unknown. In order to help resolve this critical question, we generated human iPSCs from a common fibroblast cell source using either the Yamanaka factors (OCT4, SOX2, KLF4 and cMYC) or the Thomson factors (OCT4, SOX2, NANOG and LIN28), and determined their genome-wide DNA methylation profiles. In addition to shared DNA methylation aberrations present in all our iPSCs, we identified Yamanaka-iPSC (Y-iPSC)-specific and Thomson-iPSC (T-iPSC)-specific recurrent aberrations. Strikingly, not only were the genomic locations of the aberrations different but also their types: reprogramming with Yamanaka factors mainly resulted in failure to demethylate CpGs, whereas reprogramming with Thomson factors mainly resulted in failure to methylate CpGs. Differences in the level of transcripts encoding DNMT3b and TET3 between Y-iPSCs and T-iPSCs may contribute partially to the distinct types of aberrations. Finally, <em>de novo</em> aberrantly methylated genes in Y-iPSCs were enriched for NANOG targets that are also aberrantly methylated in some cancers. Our study thus reveals that the choice of reprogramming factors influences the amount, location, and class of DNA methylation aberrations in iPSCs. These findings may provide clues into how to produce human iPSCs with fewer DNA methylation abnormalities.</p></div>","PeriodicalId":9811,"journal":{"name":"Cell Regeneration","volume":"3 1","pages":"Article 3:4"},"PeriodicalIF":0.0,"publicationDate":"2014-01-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://sci-hub-pdf.com/10.1186/2045-9769-3-4","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"32824475","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Exploring the utility of organo-polyoxometalate hybrids to inhibit SOX transcription factors","authors":"Kamesh Narasimhan ,&nbsp;Kevin Micoine ,&nbsp;Emmanuel Lacôte ,&nbsp;Serge Thorimbert ,&nbsp;Edwin Cheung ,&nbsp;Bernold Hasenknopf ,&nbsp;Ralf Jauch","doi":"10.1186/2045-9769-3-10","DOIUrl":"10.1186/2045-9769-3-10","url":null,"abstract":"<div><h3>Background</h3><p>SOX transcription factors constitute an attractive target class for intervention with small molecules as they play a prominent role in the field of regenerative biomedicine and cancer biology. However, rationally engineering specific inhibitors that interfere with transcription factor DNA interfaces continues to be a monumental challenge in the field of transcription factor chemical biology. Polyoxometalates (POMs) are inorganic compounds that were previously shown to target the high-mobility group (HMG) of SOX proteins at nanomolar concentrations. In continuation of this work, we carried out an assessment of the selectivity of a panel of newly synthesized organo-polyoxometalate hybrids in targeting different transcription factor families to enable the usage of polyoxometalates as specific SOX transcription factor drugs.</p></div><div><h3>Results</h3><p>The residual DNA-binding activities of 15 different transcription factors were measured after treatment with a panel of diverse polyoxometalates. Polyoxometalates belonging to the Dawson structural class were found to be more potent inhibitors than the Keggin class. Further, organically modified Dawson polyoxometalates were found to be the most potent in inhibiting transcription factor DNA binding activity. The size of the polyoxometalates and its derivitization were found to be the key determinants of their potency.</p></div><div><h3>Conclusion</h3><p>Polyoxometalates are highly potent, nanomolar range inhibitors of the DNA binding activity of the Sox-HMG family. However, binding assays involving a limited subset of structurally diverse polyoxometalates revealed a low selectivity profile against different transcription factor families. Further progress in achieving selectivity and deciphering structure-activity relationship of POMs require the identification of POM binding sites on transcription factors using elaborate approaches like X-ray crystallography and multidimensional NMR. In summary, our report reaffirms that transcription factors are challenging molecular architectures and that future polyoxometalate chemistry must consider further modification strategies, to address the substantial challenges involved in achieving target selectivity.</p></div>","PeriodicalId":9811,"journal":{"name":"Cell Regeneration","volume":"3 1","pages":"Article 3:10"},"PeriodicalIF":0.0,"publicationDate":"2014-01-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://sci-hub-pdf.com/10.1186/2045-9769-3-10","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"33377331","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"OCT4: A penetrant pluripotency inducer","authors":"Xuecong Wang ,&nbsp;Ralf Jauch","doi":"10.1186/2045-9769-3-6","DOIUrl":"10.1186/2045-9769-3-6","url":null,"abstract":"<div><p>Native OCT4 protein has the intrinsic ability of crossing cellular membranes to enter cells. This finding could revive efforts to induce pluripotency with proteins replacing nucleic acid-based approaches, and raises the intriguing question as to whether OCT4 can act non-cell-autonomously.</p></div>","PeriodicalId":9811,"journal":{"name":"Cell Regeneration","volume":"3 1","pages":"Article 3:6"},"PeriodicalIF":0.0,"publicationDate":"2014-01-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://sci-hub-pdf.com/10.1186/2045-9769-3-6","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"32824477","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Bioengineering of a human whole tooth: progress and challenge","authors":"Yanding Zhang ,&nbsp;YiPing Chen","doi":"10.1186/2045-9769-3-8","DOIUrl":"10.1186/2045-9769-3-8","url":null,"abstract":"<div><p>A major challenge in stem cell-based bioengineering of an implantable human tooth is to identify appropriate sources of postnatal stem cells that are odontogenic competent as the epithelial component due to the lack of enamel epithelial cells in adult teeth. In a recent issue (2013, 2:6) of <em>Cell Regeneration</em>, Cai and colleagues reported that epithelial sheets derived from human induced pluripotent stem cells (iPSCs) can functionally substitute for tooth germ epithelium to regenerate tooth-like structures, providing an appealing stem cell source for future human tooth regeneration.</p></div>","PeriodicalId":9811,"journal":{"name":"Cell Regeneration","volume":"3 1","pages":"Article 3:8"},"PeriodicalIF":0.0,"publicationDate":"2014-01-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://sci-hub-pdf.com/10.1186/2045-9769-3-8","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"32824478","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Production of transgenic pigs over-expressing the antiviral gene Mx1","authors":"Quanmei Yan ,&nbsp;Huaqiang Yang ,&nbsp;Dongshan Yang ,&nbsp;Bentian Zhao ,&nbsp;Zhen Ouyang ,&nbsp;Zhaoming Liu ,&nbsp;Nana Fan ,&nbsp;Hongsheng Ouyang ,&nbsp;Weiwang Gu ,&nbsp;Liangxue Lai","doi":"10.1186/2045-9769-3-11","DOIUrl":"10.1186/2045-9769-3-11","url":null,"abstract":"<div><p>The myxovirus resistance gene (Mx1) has a broad spectrum of antiviral activities. It is therefore an interesting candidate gene to improve disease resistance in farm animals. In this study, we report the use of somatic cell nuclear transfer (SCNT) to produce transgenic pigs over-expressing the Mx1 gene. These transgenic pigs express approximately 15–25 times more Mx1 mRNA than non-transgenic pigs, and the protein level of Mx1 was also markedly enhanced. We challenged fibroblast cells isolated from the ear skin of transgenic and control pigs with influenza A virus and classical swine fever virus (CFSV). Indirect immunofluorescence assay (IFA) revealed a profound decrease of influenza A proliferation in Mx1 transgenic cells. Growth kinetics showed an approximately 10-fold reduction of viral copies in the transgenic cells compared to non-transgenic controls. Additionally, we found that the Mx1 transgenic cells were more resistant to CSFV infection in comparison to non-transgenic cells. These results demonstrate that the Mx1 transgene can protect against viral infection in cells of transgenic pigs and indicate that the Mx1 transgene can be harnessed to develop disease-resistant pigs.</p></div>","PeriodicalId":9811,"journal":{"name":"Cell Regeneration","volume":"3 1","pages":"Article 3:11"},"PeriodicalIF":0.0,"publicationDate":"2014-01-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://sci-hub-pdf.com/10.1186/2045-9769-3-11","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"32824481","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"SNX16 negatively regulates the migration and tumorigenesis of MCF-7 cells","authors":"Leilei Zhang ,&nbsp;Dajiang Qin ,&nbsp;Chunfang Hao ,&nbsp;Xiaodong Shu ,&nbsp;Duanqing Pei","doi":"10.1186/2045-9769-2-3","DOIUrl":"10.1186/2045-9769-2-3","url":null,"abstract":"<div><h3>Background</h3><p>Sorting nexins are a large family of proteins that are associated with various components of the endosome system and they play many roles in processes such as endocytosis, intracellular protein trafficking and cell signaling. The subcellular distribution patterns of many of them remain controversial and their <em>in vivo</em> functions have not been characterized yet.</p></div><div><h3>Results</h3><p>We investigated the subcellular distribution and function of SNX16 in this study. SNX16 is detected on Rab5-positive endosomes localized adjacent to focal adhesions at cell cortex. Inhibition of SNX23, polymerization of microtubule filaments as well as the PI3-kinase all disrupt the cell cortex distribution of SNX16. Ectopic expression of SNX16 reduces the migration and the tumor formation activity of MCF-7 cells.</p></div><div><h3>Conclusion</h3><p>Our results indicate that, in addition to the PI3P, there is a SNX23- and microtubule-dependent cargo transport pathway required for the proper subcellular distribution of SNX16. SNX16 plays a negative regulatory role during cell migration and tumorigenesis.</p></div>","PeriodicalId":9811,"journal":{"name":"Cell Regeneration","volume":"2 1","pages":"Article 2:3"},"PeriodicalIF":0.0,"publicationDate":"2013-01-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://sci-hub-pdf.com/10.1186/2045-9769-2-3","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"32824078","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}