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https://www.readbyqxmd.com/read/29440377/efficient-derivation-of-stable-primed-pluripotent-embryonic-stem-cells-from-bovine-blastocysts
#1
Yanina Soledad Bogliotti, Jun Wu, Marcela Vilarino, Daiji Okamura, Delia Alba Soto, Cuiqing Zhong, Masahiro Sakurai, Rafael Vilar Sampaio, Keiichiro Suzuki, Juan Carlos Izpisua Belmonte, Pablo Juan Ross
Embryonic stem cells (ESCs) are derived from the inner cell mass of preimplantation blastocysts. From agricultural and biomedical perspectives, the derivation of stable ESCs from domestic ungulates is important for genomic testing and selection, genome engineering, and modeling human diseases. Cattle are one of the most important domestic ungulates that are commonly used for food and bioreactors. To date, however, it remains a challenge to produce stable pluripotent bovine ESC lines. Employing a culture system containing fibroblast growth factor 2 and an inhibitor of the canonical Wnt-signaling pathway, we derived pluripotent bovine ESCs (bESCs) with stable morphology, transcriptome, karyotype, population-doubling time, pluripotency marker gene expression, and epigenetic features...
February 9, 2018: Proceedings of the National Academy of Sciences of the United States of America
https://www.readbyqxmd.com/read/29301845/elixir-of-life-thwarting-aging-with-regenerative-reprogramming
#2
REVIEW
Ergin Beyret, Paloma Martinez Redondo, Aida Platero Luengo, Juan Carlos Izpisua Belmonte
All living beings undergo systemic physiological decline after ontogeny, characterized as aging. Modern medicine has increased the life expectancy, yet this has created an aged society that has more predisposition to degenerative disorders. Therefore, novel interventions that aim to extend the healthspan in parallel to the life span are needed. Regeneration ability of living beings maintains their biological integrity and thus is the major leverage against aging. However, mammalian regeneration capacity is low and further declines during aging...
January 5, 2018: Circulation Research
https://www.readbyqxmd.com/read/29224783/in%C3%A2-vivo-target-gene-activation-via-crispr-cas9-mediated-trans-epigenetic-modulation
#3
Hsin-Kai Liao, Fumiyuki Hatanaka, Toshikazu Araoka, Pradeep Reddy, Min-Zu Wu, Yinghui Sui, Takayoshi Yamauchi, Masahiro Sakurai, David D O'Keefe, Estrella Núñez-Delicado, Pedro Guillen, Josep M Campistol, Cheng-Jang Wu, Li-Fan Lu, Concepcion Rodriguez Esteban, Juan Carlos Izpisua Belmonte
Current genome-editing systems generally rely on inducing DNA double-strand breaks (DSBs). This may limit their utility in clinical therapies, as unwanted mutations caused by DSBs can have deleterious effects. CRISPR/Cas9 system has recently been repurposed to enable target gene activation, allowing regulation of endogenous gene expression without creating DSBs. However, in vivo implementation of this gain-of-function system has proven difficult. Here, we report a robust system for in vivo activation of endogenous target genes through trans-epigenetic remodeling...
November 30, 2017: Cell
https://www.readbyqxmd.com/read/29215090/in-vivo-genome-editing-via-the-hiti-method-as-a-tool-for-gene-therapy
#4
REVIEW
Keiichiro Suzuki, Juan Carlos Izpisua Belmonte
Using genome-editing technologies to correct specific mutations represents a potentially transformative new approach for treating genetic disorders. Despite rapid advances in the field of genome editing, it is still unclear whether the long-standing goal of in vivo targeted transgene integration is feasible. This is primarily because current tools are inefficient. In particular, current technologies are incapable of targeted gene knock-in in non-dividing cells, the major building blocks of adult tissues. This poses a significant barrier for developing therapeutic strategies to treat a broad range of devastating genetic disorders...
November 13, 2017: Journal of Human Genetics
https://www.readbyqxmd.com/read/28985523/human-embryo-editing-opportunities-and-importance-of-transnational-cooperation
#5
Duanqing Pei, David W Beier, Ephrat Levy-Lahad, Gary Marchant, Janet Rossant, Juan Carlos Izpisua Belmonte, Robin Lovell-Badge, Rudolf Jaenisch, Alta Charo, David Baltimore
A recent National Academies report articulates a path forward for research, ethics, and governance of clinical applications involving genome editing. In light of recent human embryo editing developments, scientists and stakeholders from all nations should cooperate to take advantage of this historic opportunity for medicine and also basic human biology.
October 5, 2017: Cell Stem Cell
https://www.readbyqxmd.com/read/28920955/mir-25-93-mediates-hypoxia-induced-immunosuppression-by-repressing-cgas
#6
Min-Zu Wu, Wei-Chung Cheng, Su-Feng Chen, Shin Nieh, Carolyn O'Connor, Chia-Lin Liu, Wen-Wei Tsai, Cheng-Jang Wu, Lorena Martin, Yaoh-Shiang Lin, Kou-Juey Wu, Li-Fan Lu, Juan Carlos Izpisua Belmonte
The mechanisms by which hypoxic tumours evade immunological pressure and anti-tumour immunity remain elusive. Here, we report that two hypoxia-responsive microRNAs, miR-25 and miR-93, are important for establishing an immunosuppressive tumour microenvironment by downregulating expression of the DNA sensor cGAS. Mechanistically, miR-25/93 targets NCOA3, an epigenetic factor that maintains basal levels of cGAS expression, leading to repression of cGAS during hypoxia. This allows hypoxic tumour cells to escape immunological responses induced by damage-associated molecular pattern molecules, specifically the release of mitochondrial DNA...
October 2017: Nature Cell Biology
https://www.readbyqxmd.com/read/28888991/lrrk2-functions-as-a-scaffolding-kinase-of-ask1-mediated-neuronal-cell-death
#7
Ji-Hye Yoon, Jung-Soon Mo, Mi-Yeon Kim, Eun-Jung Ann, Ji-Seon Ahn, Eun-Hye Jo, Hye-Jin Lee, Young Chul Lee, Wongi Seol, Sergiy M Yarmoluk, Thomas Gasser, Philipp J Kahle, Guang-Hui Liu, Juan Carlos Izpisua Belmonte, Hee-Sae Park
Leucine-rich repeat kinase 2 (LRRK2), a multi-domain protein, is a key causative factor in Parkinson's disease (PD). Identification of novel substrates and the molecular mechanisms underlying the effects of LRRK2 are essential for understanding the pathogenesis of PD. In this study, we showed that LRRK2 played an important role in neuronal cell death by directly phosphorylating and activating apoptosis signal-regulating kinase 1 (ASK1). LRRK2 phosphorylated ASK1 at Thr832 that is adjacent to Thr845, which serves as an autophosphorylation site...
December 2017: Biochimica et Biophysica Acta
https://www.readbyqxmd.com/read/28874671/crispr-cas9-mediated-one-step-disabling-of-pancreatogenesis-in-pigs
#8
Jun Wu, Marcela Vilarino, Keiichiro Suzuki, Daiji Okamura, Yanina Soledad Bogliotti, Insung Park, Joan Rowe, Bret McNabb, Pablo Juan Ross, Juan Carlos Izpisua Belmonte
Genome editing using programmable nucleases has revolutionized biomedical research. CRISPR-Cas9 mediated zygote genome editing enables high efficient production of knockout animals suitable for studying development and relevant human diseases. Here we report efficient disabling pancreatogenesis in pig embryos via zygotic co-delivery of Cas9 mRNA and dual sgRNAs targeting the PDX1 gene, which when combined with chimeric-competent human pluriopotent stem cells may serve as a suitable platform for the xeno-generation of human tissues and organs in pigs...
September 5, 2017: Scientific Reports
https://www.readbyqxmd.com/read/28826795/regulation-of-stem-cell-aging-by-metabolism-and-epigenetics
#9
REVIEW
Ruotong Ren, Alejandro Ocampo, Guang-Hui Liu, Juan Carlos Izpisua Belmonte
Stem cell aging and exhaustion are considered important drivers of organismal aging. Age-associated declines in stem cell function are characterized by metabolic and epigenetic changes. Understanding the mechanisms underlying these changes will likely reveal novel therapeutic targets for ameliorating age-associated phenotypes and for prolonging human healthspan. Recent studies have shown that metabolism plays an important role in regulating epigenetic modifications and that this regulation dramatically affects the aging process...
September 5, 2017: Cell Metabolism
https://www.readbyqxmd.com/read/28802034/keeping-the-rhythm-while-changing-the-lyrics-circadian-biology-in-aging
#10
COMMENT
Fumiyuki Hatanaka, Alejandro Ocampo, Juan Carlos Izpisua Belmonte
Aging and circadian rhythms have been linked for decades, but their molecular interplay has remained obscure. Sato et al. and Solanas et al. now reveal that, while core clock components remain nearly unaltered, aging is associated with tissue-specific rewiring, which can be prevented by calorie restriction.
August 10, 2017: Cell
https://www.readbyqxmd.com/read/28783728/correction-of-a-pathogenic-gene-mutation-in-human-embryos
#11
Hong Ma, Nuria Marti-Gutierrez, Sang-Wook Park, Jun Wu, Yeonmi Lee, Keiichiro Suzuki, Amy Koski, Dongmei Ji, Tomonari Hayama, Riffat Ahmed, Hayley Darby, Crystal Van Dyken, Ying Li, Eunju Kang, A-Reum Park, Daesik Kim, Sang-Tae Kim, Jianhui Gong, Ying Gu, Xun Xu, David Battaglia, Sacha A Krieg, David M Lee, Diana H Wu, Don P Wolf, Stephen B Heitner, Juan Carlos Izpisua Belmonte, Paula Amato, Jin-Soo Kim, Sanjiv Kaul, Shoukhrat Mitalipov
Genome editing has potential for the targeted correction of germline mutations. Here we describe the correction of the heterozygous MYBPC3 mutation in human preimplantation embryos with precise CRISPR-Cas9-based targeting accuracy and high homology-directed repair efficiency by activating an endogenous, germline-specific DNA repair response. Induced double-strand breaks (DSBs) at the mutant paternal allele were predominantly repaired using the homologous wild-type maternal gene instead of a synthetic DNA template...
August 24, 2017: Nature
https://www.readbyqxmd.com/read/28728561/understanding-the-genetics-behind-complex-human-disease-with-large-scale-ipsc-collections
#12
Amanda E Yamasaki, Athanasia D Panopoulos, Juan Carlos Izpisua Belmonte
Three recent studies analyzing large-scale collections of human induced pluripotent stem cell lines provide valuable insight into how genetic regulatory variation affects cellular and molecular traits.
July 20, 2017: Genome Biology
https://www.readbyqxmd.com/read/28685772/genetic-enhancement-in-cultured-human-adult-stem-cells-conferred-by-a-single-nucleotide-recoding
#13
Jiping Yang, Jingyi Li, Keiichiro Suzuki, Xiaomeng Liu, Jun Wu, Weiqi Zhang, Ruotong Ren, Weizhou Zhang, Piu Chan, Juan Carlos Izpisua Belmonte, Jing Qu, Fuchou Tang, Guang-Hui Liu
No abstract text is available yet for this article.
September 2017: Cell Research
https://www.readbyqxmd.com/read/28586330/gathering-by-the-red-sea-highlights-links-between-environment-and-epigenetics
#14
Mo Li, Emiliana Borrelli, Pierre J Magistretti, Juan Carlos Izpisua Belmonte, Paolo Sassone-Corsi, Valerio Orlando
No abstract text is available yet for this article.
June 6, 2017: Nature Structural & Molecular Biology
https://www.readbyqxmd.com/read/28473583/integration-of-cpg-free-dna-induces-de-novo-methylation-of-cpg-islands-in-pluripotent-stem-cells
#15
Yuta Takahashi, Jun Wu, Keiichiro Suzuki, Paloma Martinez-Redondo, Mo Li, Hsin-Kai Liao, Min-Zu Wu, Reyna Hernández-Benítez, Tomoaki Hishida, Maxim Nikolaievich Shokhirev, Concepcion Rodriguez Esteban, Ignacio Sancho-Martinez, Juan Carlos Izpisua Belmonte
CpG islands (CGIs) are primarily promoter-associated genomic regions and are mostly unmethylated within highly methylated mammalian genomes. The mechanisms by which CGIs are protected from de novo methylation remain elusive. Here we show that insertion of CpG-free DNA into targeted CGIs induces de novo methylation of the entire CGI in human pluripotent stem cells (PSCs). The methylation status is stably maintained even after CpG-free DNA removal, extensive passaging, and differentiation. By targeting the DNA mismatch repair gene MLH1 CGI, we could generate a PSC model of a cancer-related epimutation...
May 5, 2017: Science
https://www.readbyqxmd.com/read/28467430/analysis-of-transcription-factors-expressed-at-the-anterior-mouse-limb-bud
#16
Shigetoshi Yokoyama, Soichi Furukawa, Shoya Kitada, Masaki Mori, Takeshi Saito, Koichi Kawakami, Juan Carlos Izpisua Belmonte, Yasuhiko Kawakami, Yoshiaki Ito, Tempei Sato, Hiroshi Asahara
Limb bud patterning, outgrowth, and differentiation are precisely regulated in a spatio-temporal manner through integrated networks of transcription factors, signaling molecules, and downstream genes. However, the exact mechanisms that orchestrate morphogenesis of the limb remain to be elucidated. Previously, we have established EMBRYS, a whole-mount in situ hybridization database of transcription factors. Based on the findings from EMBRYS, we focused our expression pattern analysis on a selection of transcription factor genes that exhibit spatially localized and temporally dynamic expression patterns with respect to the anterior-posterior axis in the E9...
2017: PloS One
https://www.readbyqxmd.com/read/28455313/transient-transcription-factor-oskm-expression-is-key-towards-clinical-translation-of-in%C3%A2-vivo-cell-reprogramming
#17
Irene de Lázaro, Giulio Cossu, Kostas Kostarelos
Reprogramming adult, fully differentiated cells to pluripotency in vivo via Oct3/4, Sox2, Klf4 and c-Myc (OSKM) overexpression has proved feasible in various independent studies and could be used to induce tissue regeneration owing to the proliferative capacity and differentiation potential of the reprogrammed cells. However, a number of these reports have described the generation of teratomas caused by sustained reprogramming, which precludes the therapeutic translation of this technology. A recent study by the Izpisúa-Belmonte laboratory described a cyclic regime for short-term OSKM expression in vivo that prevents complete reprogramming to the pluripotent state as well as tumorigenesis...
June 2017: EMBO Molecular Medicine
https://www.readbyqxmd.com/read/28410642/ipscore-a-resource-of-222-ipsc-lines-enabling-functional-characterization-of-genetic-variation-across-a-variety-of-cell-types
#18
Athanasia D Panopoulos, Matteo D'Antonio, Paola Benaglio, Roy Williams, Sherin I Hashem, Bernhard M Schuldt, Christopher DeBoever, Angelo D Arias, Melvin Garcia, Bradley C Nelson, Olivier Harismendy, David A Jakubosky, Margaret K R Donovan, William W Greenwald, KathyJean Farnam, Megan Cook, Victor Borja, Carl A Miller, Jonathan D Grinstein, Frauke Drees, Jonathan Okubo, Kenneth E Diffenderfer, Yuriko Hishida, Veronica Modesto, Carl T Dargitz, Rachel Feiring, Chang Zhao, Aitor Aguirre, Thomas J McGarry, Hiroko Matsui, He Li, Joaquin Reyna, Fangwen Rao, Daniel T O'Connor, Gene W Yeo, Sylvia M Evans, Neil C Chi, Kristen Jepsen, Naoki Nariai, Franz-Josef Müller, Lawrence S B Goldstein, Juan Carlos Izpisua Belmonte, Eric Adler, Jeanne F Loring, W Travis Berggren, Agnieszka D'Antonio-Chronowska, Erin N Smith, Kelly A Frazer
Large-scale collections of induced pluripotent stem cells (iPSCs) could serve as powerful model systems for examining how genetic variation affects biology and disease. Here we describe the iPSCORE resource: a collection of systematically derived and characterized iPSC lines from 222 ethnically diverse individuals that allows for both familial and association-based genetic studies. iPSCORE lines are pluripotent with high genomic integrity (no or low numbers of somatic copy-number variants) as determined using high-throughput RNA-sequencing and genotyping arrays, respectively...
April 11, 2017: Stem Cell Reports
https://www.readbyqxmd.com/read/28401346/crispr-cas9-mediated-targeted-gene-correction-in-amyotrophic-lateral-sclerosis-patient-ipscs
#19
Lixia Wang, Fei Yi, Lina Fu, Jiping Yang, Si Wang, Zhaoxia Wang, Keiichiro Suzuki, Liang Sun, Xiuling Xu, Yang Yu, Jie Qiao, Juan Carlos Izpisua Belmonte, Ze Yang, Yun Yuan, Jing Qu, Guang-Hui Liu
Amyotrophic lateral sclerosis (ALS) is a complex neurodegenerative disease with cellular and molecular mechanisms yet to be fully described. Mutations in a number of genes including SOD1 and FUS are associated with familial ALS. Here we report the generation of induced pluripotent stem cells (iPSCs) from fibroblasts of familial ALS patients bearing SOD1 (+/A272C) and FUS (+/G1566A) mutations, respectively. We further generated gene corrected ALS iPSCs using CRISPR/Cas9 system. Genome-wide RNA sequencing (RNA-seq) analysis of motor neurons derived from SOD1 (+/A272C) and corrected iPSCs revealed 899 aberrant transcripts...
May 2017: Protein & Cell
https://www.readbyqxmd.com/read/28392216/ipscore-a-resource-of-222-ipsc-lines-enabling-functional-characterization-of-genetic-variation-across-a-variety-of-cell-types
#20
Athanasia D Panopoulos, Matteo D'Antonio, Paola Benaglio, Roy Williams, Sherin I Hashem, Bernhard M Schuldt, Christopher DeBoever, Angelo D Arias, Melvin Garcia, Bradley C Nelson, Olivier Harismendy, David A Jakubosky, Margaret K R Donovan, William W Greenwald, KathyJean Farnam, Megan Cook, Victor Borja, Carl A Miller, Jonathan D Grinstein, Frauke Drees, Jonathan Okubo, Kenneth E Diffenderfer, Yuriko Hishida, Veronica Modesto, Carl T Dargitz, Rachel Feiring, Chang Zhao, Aitor Aguirre, Thomas J McGarry, Hiroko Matsui, He Li, Joaquin Reyna, Fangwen Rao, Daniel T O'Connor, Gene W Yeo, Sylvia M Evans, Neil C Chi, Kristen Jepsen, Naoki Nariai, Franz-Josef Müller, Lawrence S B Goldstein, Juan Carlos Izpisua Belmonte, Eric Adler, Jeanne F Loring, W Travis Berggren, Agnieszka D'Antonio-Chronowska, Erin N Smith, Kelly A Frazer
Large-scale collections of induced pluripotent stem cells (iPSCs) could serve as powerful model systems for examining how genetic variation affects biology and disease. Here we describe the iPSCORE resource: a collection of systematically derived and characterized iPSC lines from 222 ethnically diverse individuals that allows for both familial and association-based genetic studies. iPSCORE lines are pluripotent with high genomic integrity (no or low numbers of somatic copy-number variants) as determined using high-throughput RNA-sequencing and genotyping arrays, respectively...
April 3, 2017: Stem Cell Reports
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