Altered A-to-I RNA Editing in Human Embryogenesis
Ronit Shtrichman,Igal Germanguz,Rachel Mandel,Anna Ziskind,Irit Nahor,Michal Safran,Sivan Osenberg,Ofra Sherf,Gideon Rechavi,Joseph Itskovitz-Eldor +9 more
TLDR
The data suggest that A-to-I RNA editing plays a critical role during early human development, and suggests that ADAR1 protein is substantially regulated in undifferentiated pluripotent hESCs.Abstract:
Post-transcriptional events play an important role in human development. The question arises as to whether Adenosine to Inosine RNA editing, catalyzed by the ADAR (Adenosine Deaminase acting on RNA) enzymes, differs in human embryogenesis and in adulthood. We tested the editing of various target genes in coding (FLNA, BLCAP, CYFIP2) and non-coding sequences at their Alu elements (BRCA1, CARD11, RBBP9, MDM4, FNACC), as well as the transcriptional levels of the ADAR1 enzymes. This analysis was performed on five fetal and adult human tissues: brain, heart, liver, kidney, and spleen, as well as on human embryonic stem cells (hESCs), which represent the blastocyst stage in early human development. Our results show substantially greater editing activity for most adult tissue samples relative to fetal ones, in six of the eight genes tested. To test the effect of reduced A-to-I RNA editing activity in early human development we used human embryonic stem cells (hESCs) as a model and tried to generate hESC clones that overexpress the ADAR1-p110 isoform. We were unable to achieve overexpression of ADAR1-p110 by either transfection or lentiviral infection, though we easily generated hESC clones that expressed the GFP transgene and overexpressed ADAR1-p110 in 293T cells and in primary human foreskin fibroblast (HFF) cells. Moreover, in contrast to the expected overexpression of ADAR1-p110 protein following its introduction into hESCs, the expression levels of this protein decreased dramatically 24-48 hr post infection. Similar results were obtained when we tried to overexpress ADAR1-p110 in pluripotent embryonal carcinoma cells. This suggests that ADAR1 protein is substantially regulated in undifferentiated pluripotent hESCs. Overall, our data suggest that A-to-I RNA editing plays a critical role during early human development.read more
Citations
More filters
Journal ArticleDOI
Circular RNAs in the Mammalian Brain Are Highly Abundant, Conserved, and Dynamically Expressed
Agnieszka Rybak-Wolf,Christin Stottmeister,Petar Glažar,Marvin Jens,Natalia S. Pino,Sebastian A. Giusti,Mor Hanan,Mikaela Behm,Osnat Bartok,Reut Ashwal-Fluss,Margareta Herzog,Luisa Schreyer,Panagiotis Papavasileiou,Andranik Ivanov,Marie Öhman,Damian Refojo,Sebastian Kadener,Nikolaus Rajewsky +17 more
TL;DR: A circRNA brain expression atlas and evidence for important circRNA functions and values as biomarkers are provided and discovered and analyzed thousands of neuronal human and mouse circRNAs.
Journal ArticleDOI
Adenosine Deaminase That Acts on RNA 3 (ADAR3) Binding to Glutamate Receptor Subunit B Pre-mRNA Inhibits RNA Editing in Glioblastoma
TL;DR: This study identifies adenosine deaminase that acts on RNA 3 (ADAR3) as an important regulator of Q/R site editing, investigates its mode of action, and detects elevated ADAR3 expression in glioblastoma tumors compared with adjacent brain tissue.
Journal ArticleDOI
Circular RNAs: Novel Regulators of Neuronal Development.
TL;DR: Evidence from analysis of some circRNA molecules suggests that they could substantially contribute to the regulation of gene expression, particularly in architecturally complex and polarized cells such as neurons.
Journal ArticleDOI
To edit or not to edit: regulation of ADAR editing specificity and efficiency.
TL;DR: During normal development, in specific cells, and in both neuropathological diseases and cancers, the extent of RNA editing does not directly correlate with levels of the substrate mRNA or the adenosine deaminase that act on RNA (ADAR) editing enzymes, implying that cellular factors are required for spatiotemporal regulation of A‐to‐I editing.
Journal ArticleDOI
The cultural divide: exponential growth in classical 2D and metabolic equilibrium in 3D environments.
Krzysztof Wrzesinski,Adelina Rogowska-Wrzesinska,Rattiyaporn Kanlaya,Kamil Borkowski,Veit Schwämmle,Jie Dai,Kira Joensen,Katarzyna Wojdyla,Vasco Botelho Carvalho,Stephen J. Fey +9 more
TL;DR: It is demonstrated that cells at dynamic equilibrium can match the physiological performance of cells in tissues in vivo and the multitude of protein changes necessary to achieve this performance are described.
References
More filters
Journal ArticleDOI
Embryonic Stem Cell Lines Derived from Human Blastocysts
James A. Thomson,Joseph Itskovitz-Eldor,Sander S. Shapiro,Michelle A. Waknitz,Swiergiel Jennifer J,Vivienne S. Marshall,Jeffrey M. Jones +6 more
TL;DR: Human blastocyst-derived, pluripotent cell lines are described that have normal karyotypes, express high levels of telomerase activity, and express cell surface markers that characterize primate embryonic stem cells but do not characterize other early lineages.
Journal ArticleDOI
Stem cells, cancer, and cancer stem cells
TL;DR: Stem cell biology has come of age: Unequivocal proof that stem cells exist in the haematopoietic system has given way to the prospective isolation of several tissue-specific stem and progenitor cells, the initial delineation of their properties and expressed genetic programmes, and the beginnings of their utility in regenerative medicine.
Journal ArticleDOI
A gene atlas of the mouse and human protein-encoding transcriptomes
Andrew I. Su,Tim Wiltshire,Serge Batalov,Hilmar Lapp,Keith A. Ching,David Block,Jie Zhang,Richard Soden,Mimi Hayakawa,Gabriel Kreiman,Gabriel Kreiman,Michael P. Cooke,John R. Walker,John B. Hogenesch,John B. Hogenesch +14 more
TL;DR: In this paper, high-density oligonucleotide arrays offer the opportunity to examine patterns of gene expression on a genome scale, and the authors have designed custom arrays that interrogate the expression of the vast majority of proteinencoding human and mouse genes and have used them to profile a panel of 79 human and 61 mouse tissues.
Journal ArticleDOI
Clonally Derived Human Embryonic Stem Cell Lines Maintain Pluripotency and Proliferative Potential for Prolonged Periods of Culture
Michal Amit,Melissa K. Carpenter,Margaret S. Inokuma,Choy-Pik Chiu,Charles P. Harris,Michelle A. Waknitz,Joseph Itskovitz-Eldor,James A. Thomson +7 more
TL;DR: The clonal derivation of two human ES cell lines, H9.1 and H.2, demonstrates the pluripotency of single human ES cells, the maintenance of pluripOTency during an extended period of culture, and the long-term self-renewing properties of cultured human ES Cells.
Journal ArticleDOI
RNA Editing by Adenosine Deaminases That Act on RNA
TL;DR: This review summarizes ongoing investigations of the enzyme family and their substrates, focusing on biological function as well as biochemical mechanism.