Tissue-specific and ubiquitous gene knockouts by TALEN electroporation provide new approaches to investigating gene function in Ciona

Development
Nicholas TreenYasunori Sasakura

Abstract

Custom designed nucleases can simplify gene targeting experiments and have the potential to allow these techniques to be performed in a wide range of organisms. Transcriptional activator-like effector nucleases (TALENs) are starting to fulfill this potential with the advantages of low cost and fast construction times. Here, we report that TALENs are highly effective at inducing mutations in specific genomic loci in the ascidian chordate Ciona intestinalis. In Ciona there are well-established methods to introduce exogenous DNA by electroporation, and we show that this method can be used to introduce constructs that can express TALENs ubiquitously or in specific tissues. Our current protocols enable the rapid analysis of hundreds of TALEN-induced mutants. TALEN electroporations result in a high rate of mutations. These mutations can result in gene knockouts that recapitulate previously described functions of Fgf3 and Hox12. We show that TALENs can work efficiently to cause tissue-specific knockouts and demonstrate this by knocking out Hox12 in the epidermis and Fgf3 in neural tissues. We also use tissue-specific knockouts to reveal a new function of Fgf3 during ascidian larval metamorphosis.

References

Jun 16, 1989·Science·M R Capecchi
Sep 27, 2002·Genesis : the Journal of Genetics and Development·Joseph B Duffy
Oct 10, 2002·Development Genes and Evolution·Yutaka SatouNori Satoh
Jun 6, 2003·Proceedings of the National Academy of Sciences of the United States of America·Yasunori SasakuraNori Satoh
Aug 6, 2005·Nature Biotechnology·Matthew H Porteus, Dana Carroll
May 27, 2006·Science·Kaoru S ImaiYutaka Satou
Mar 17, 2007·Developmental Dynamics : an Official Publication of the American Association of Anatomists·Gaku Kumano, Hiroki Nishida
Oct 26, 2007·Developmental Biology·Matthew J Kourakis, William C Smith
Apr 12, 2008·Development·Judith S Eisen, James C Smith
May 27, 2008·Nature Biotechnology·Xiangdong MengScot A Wolfe
Jun 3, 2008·Trends in Genetics : TIG·Arhat AbzhanovAntonia Monteiro
Nov 21, 2008·Development·Shota ChibaWilliam C Smith
Dec 23, 2008·Developmental Biology·Akie Nakayama-IshimuraYasunori Sasakura
Dec 24, 2010·Nature Biotechnology·Jeffrey C MillerEdward J Rebar
May 12, 2011·Development·Patrick Lemaire
Nov 24, 2011·Development·Alberto StolfiMichael Levine
Jan 10, 2012·Nature Protocols·Neville E SanjanaFeng Zhang
Apr 10, 2012·Nature Biotechnology·Deepak ReyonJ Keith Joung
May 30, 2012·Development, Growth & Differentiation·Narudo KawaiYasunori Sasakura
Sep 25, 2012·Nature·Victoria M BedellStephen C Ekker
Oct 3, 2012·Proceedings of the National Academy of Sciences of the United States of America·Daniel F CarlsonScott C Fahrenkrug
Oct 10, 2012·Proceedings of the National Academy of Sciences of the United States of America·Yong LeiHui Zhao
Feb 8, 2013·Genes to Cells : Devoted to Molecular & Cellular Mechanisms·Tetsushi SakumaTakashi Yamamoto
Feb 19, 2013·Nature Biotechnology·Yongsub KimJin-Soo Kim
Nov 30, 2013·Scientific Reports·Tetsushi SakumaTakashi Yamamoto

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Citations

Mar 13, 2014·Annual Review of Biochemistry·Dana Carroll
Oct 1, 2015·Nucleic Acids Research·Matija BrozovicPatrick Lemaire
Aug 8, 2015·Developmental Dynamics : an Official Publication of the American Association of Anatomists·Akiko HozumiYasunori Sasakura
Oct 8, 2015·Blood Reviews·Xiangguo ShiAnskar Y H Leung
Sep 13, 2014·Development, Growth & Differentiation·Haruka SasakiYasunori Sasakura
Mar 13, 2014·Genesis : the Journal of Genetics and Development·Keita YoshidaYasunori Sasakura
Mar 31, 2015·Current Opinion in Genetics & Development·Nicole KaplanLionel Christiaen
Nov 15, 2014·Genesis : the Journal of Genetics and Development·Fabio CrocettaPaolo Sordino
Sep 25, 2014·Genesis : the Journal of Genetics and Development·Anthi KaraiskouJean-Philippe Chambon
Sep 30, 2014·Genesis : the Journal of Genetics and Development·Michael Veeman, Wendy Reeves
Jul 25, 2014·The Biochemical Journal·David A WrightMartin H Spalding
Apr 19, 2015·Developmental Biology·Narudo KawaiYasunori Sasakura
Mar 5, 2016·Genesis : the Journal of Genetics and Development·Wei Li, Guangshuo Ou
Oct 16, 2016·Marine Genomics·Tsuyoshi Momose, Jean-Paul Concordet
Dec 23, 2016·Proceedings. Biological Sciences·Yasunori SasakuraKeita Yoshida
Sep 24, 2015·Anais Da Academia Brasileira De Ciências·Maurício Rocha-MartinsRodrigo A P Martins
Jun 10, 2016·Evolutionary Bioinformatics Online·Edward C StanleySteven Q Irvine
Feb 15, 2020·Scientific Reports·Hitoshi SawadaMaki Shirae-Kurabayashi
Jun 8, 2017·Mammalian Genome : Official Journal of the International Mammalian Genome Society·Michelle N Knowlton, Cynthia L Smith
Nov 7, 2017·Wiley Interdisciplinary Reviews. Developmental Biology·Yasunori Sasakura, Akiko Hozumi
Mar 30, 2020·Current Biology : CB·Akiko HozumiYasunori Sasakura
Nov 5, 2019·Developmental Biology·Yukako TajimaYasunori Sasakura
Oct 23, 2014·Development·Ying PengStephen C Ekker
Oct 23, 2014·Development·Alberto StolfiLionel Christiaen

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