Mammalian nuclear transplantation to Germinal Vesicle stage Xenopus oocytes - a method for quantitative transcriptional reprogramming.

Methods : a Companion to Methods in Enzymology
Richard P Halley-StottJohn B Gurdon

Abstract

Full-grown Xenopus oocytes in first meiotic prophase contain an immensely enlarged nucleus, the Germinal Vesicle (GV), that can be injected with several hundred somatic cell nuclei. When the nuclei of mammalian somatic cells or cultured cell lines are injected into a GV, a wide range of genes that are not transcribed in the donor cells, including pluripotency genes, start to be transcriptionally activated, and synthesize primary transcripts continuously for several days. Because of the large size and abundance of Xenopus laevis oocytes, this experimental system offers an opportunity to understand the mechanisms by which somatic cell nuclei can be reprogrammed to transcribe genes characteristic of oocytes and early embryos. The use of mammalian nuclei ensures that there is no background of endogenous maternal transcripts of the kind that are induced. The induced gene transcription takes place in the absence of cell division or DNA synthesis and does not require protein synthesis. Here we summarize new as well as established results that characterize this experimental system. In particular, we describe optimal conditions for transplanting somatic nuclei to oocytes and for the efficient activation of transcription by transplanted ...Continue Reading

References

Dec 1, 1977·Cell·T G Clark, R W Merriam
Nov 3, 1989·Science·R A LaskeyJ J Blow
Apr 26, 1974·Nature·J B Gurdon
Mar 22, 1982·Proceedings of the Royal Society of London. Series B, Containing Papers of a Biological Character·H G Callan
Jan 10, 2003·Nucleic Acids Research·Filip PattynJo Vandesompele
Jun 1, 1958·Experimental Cell Research·T R ELSDALES SMITH
Sep 13, 2007·Stem Cell Reviews·Philippe Collas, Christel K Taranger
Feb 13, 2008·PLoS Genetics·Susana M Chuva de Sousa LopesAnne McLaren
May 15, 2009·Epigenetics : Official Journal of the DNA Methylation Society·Yuhong BianAndrew D Johnson
Dec 9, 2009·Methods : a Companion to Methods in Enzymology·Stefaan DerveauxJan Hellemans
Mar 10, 2010·Proceedings of the National Academy of Sciences of the United States of America·Jerome JullienJohn B Gurdon

❮ Previous
Next ❯

Citations

Jan 28, 2014·Nature·Maria A ChristophorouTony Kouzarides
Apr 10, 2014·Chemical Communications : Chem Comm·Michael C YoungRichard J Hooley
May 7, 2014·Differentiation; Research in Biological Diversity·Gabriele Colozza, Edward M De Robertis
Nov 5, 2010·Cold Spring Harbor Symposia on Quantitative Biology·V PasqueJ B Gurdon
Jul 30, 2014·PLoS Biology·Richard P Halley-StottJohn Gurdon
Jan 1, 2013·Cellular and Molecular Life Sciences : CMLS·Kei Miyamoto, J B Gurdon
Nov 15, 2012·Chromosome Research : an International Journal on the Molecular, Supramolecular and Evolutionary Aspects of Chromosome Biology·May Penrad-MobayedJean-Antoine Lepesant
Aug 9, 2011·Nature Biotechnology·Uta GrieshammerAlan Trounson
Apr 16, 2013·Briefings in Functional Genomics·Richard P Halley-Stott, John B Gurdon
Aug 13, 2011·Biologie aujourd'hui·Jérôme Jullien, John Gurdon
May 10, 2011·The EMBO Journal·Vincent PasqueJohn B Gurdon
Apr 11, 2012·Genes to Cells : Devoted to Molecular & Cellular Mechanisms·Maria Manukyan, Prim B Singh
Dec 7, 2013·Angewandte Chemie·John B Gurdon
Jan 7, 2015·Genes & Development·Thais LavagnolliMatthias Merkenschlager
Oct 13, 2012·Communicative & Integrative Biology·Patrick NarbonneJ B Gurdon
Sep 24, 2011·Trends in Genetics : TIG·Vincent PasqueJ B Gurdon
Oct 30, 2013·PloS One·Peng-Cheng KongMan-Xi Jiang
Dec 2, 2011·PLoS Biology·Patrick NarbonneJohn B Gurdon
May 9, 2018·Stem Cells and Development·Elena Gonzalez-Munoz, Jose B Cibelli
Jun 24, 2011·Nature Reviews. Molecular Cell Biology·Jerome JullienJ B Gurdon
Feb 13, 2019·The Journal of Reproduction and Development·Kei Miyamoto
Jun 14, 2020·Proceedings of the National Academy of Sciences of the United States of America·J B GurdonNigel Garrett
May 30, 2013·Development·J B Gurdon
May 30, 2013·Development·Richard P Halley-StottJ B Gurdon
Mar 5, 2013·Cold Spring Harbor Protocols·Yaping Lin-Moshier, Jonathan S Marchant

❮ Previous
Next ❯

Methods Mentioned

BETA
PCR
in vitro transcription

Related Concepts

Related Feeds

Aminoglycosides (ASM)

Aminoglycoside is a medicinal and bacteriologic category of traditional Gram-negative antibacterial medications that inhibit protein synthesis and contain as a portion of the molecule an amino-modified glycoside. Discover the latest research on aminoglycoside here.

Aminoglycosides

Aminoglycoside is a medicinal and bacteriologic category of traditional Gram-negative antibacterial medications that inhibit protein synthesis and contain as a portion of the molecule an amino-modified glycoside. Discover the latest research on aminoglycoside here.