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Key Message: Expression of Cre recombinase by AtRps5a pro or AtDD45 pro enabled Cre/lox-mediated recombination at an early embryonic developmental stage upon crossing, activating transgenes in the hybrid cowpea and tobacco. Genetic engineering ideally results in precise spatiotemporal control of transgene expression. To activate transgenes exclusively in a hybrid upon fertilization, we evaluated a Cre/lox-mediated gene activation system with the Cre recombinase expressed by either AtRps5a or AtDD45 promoters that showed activity in egg cells and young embryos. In crosses between Cre recombinase lines and transgenic lines harboring a lox-excision reporter cassette with ZsGreen driven by the AtUbq3 promoter after Cre/lox-mediated recombination, we observed complete excision of the lox-flanked intervening DNA sequence between the AtUbq3 pro and the ZsGreen coding sequence in F 1 progeny upon genotyping but no ZsGreen expression in F 1 seeds or seedlings. The incapability to observe ZsGreen fluorescence was attributed to the activity of the AtUbq3 pro . Strong ZsGreen expression in F 1 seeds was observed after recombination when ZsGreen was driven by the AtUbq10 promoter. Using the AtDD45 pro to express Cre resulted in more variation in recombination frequencies between transgenic lines and crosses. Regardless of the promoter used to regulate Cre, mosaic F 1 progeny were rare, suggesting gene activation at an early embryo-developmental stage. Observation of ZsGreen-expressing tobacco embryos at the globular stage from crosses with the AtRps5a pro Cre lines pollinated by the AtUbq3 pro lox line supported the early activation mode.

Titel:	Gene activation via Cre/lox-mediated excision in cowpea (Vigna unguiculata).
Autor/in / Beteiligte Person:	Zhang, Z ; Guo, Y ; Marasigan, KM ; Conner, JA ; Ozias-Akins, P
Link:	View record at Springer (Volltext)
Zeitschrift:	Plant cell reports, Jg. 41 (2022), Heft 1, S. 119-138
Veröffentlichung:	Berlin ; New York : Springer, 1981-, 2022
Medientyp:	academicJournal
ISSN:	1432-203X (electronic)
DOI:	10.1007/s00299-021-02789-z
Schlagwort:	Integrases metabolism Plant Proteins metabolism Plants, Genetically Modified Vigna enzymology Genes, Plant Integrases genetics Plant Proteins genetics Transcriptional Activation Transgenes Vigna genetics
Sonstiges:	Nachgewiesen in: MEDLINE Sprachen: English Publication Type: Journal Article Language: English [Plant Cell Rep] 2022 Jan; Vol. 41 (1), pp. 119-138. <i>Date of Electronic Publication: </i>2021 Sep 30. MeSH Terms: Genes, Plant* ; Transcriptional Activation* ; Transgenes* ; Integrases / genetics ; Plant Proteins / genetics ; Vigna / *genetics ; Integrases / metabolism ; Plant Proteins / metabolism ; Plants, Genetically Modified ; Vigna / enzymology References: Theor Appl Genet. 2003 Nov;107(7):1157-68. (PMID: 14513214) ; Plant Cell Rep. 2015 Mar;34(3):469-81. (PMID: 25504050) ; Sci Rep. 2017 Nov 6;7(1):14505. (PMID: 29109405) ; Gates Open Res. 2018 Feb 9;2:7. (PMID: 29528046) ; PLoS One. 2008;3(11):e3647. (PMID: 18985154) ; Plant J. 2021 May;106(3):817-830. (PMID: 33595147) ; Plant J. 2016 Oct;88(2):318-327. (PMID: 27448251) ; Mol Gen Genet. 1992 Jul;234(1):49-59. (PMID: 1495484) ; Front Plant Sci. 2020 Sep 16;11:567425. (PMID: 33072144) ; Plant Cell. 1990 Dec;2(12):1201-1224. (PMID: 12354953) ; Plant Mol Biol. 1992 Jan;18(2):353-61. (PMID: 1310059) ; Genes Dev. 2004 Aug 15;18(16):1964-9. (PMID: 15314023) ; Plant Cell Rep. 2011 Nov;30(11):2013-25. (PMID: 21706298) ; Plant J. 2018 Mar;93(6):1129-1142. (PMID: 29356213) ; Plant Cell Rep. 2006 Apr;25(4):304-12. (PMID: 16244884) ; Plant J. 2007 Jul;51(2):281-92. (PMID: 17559508) ; PLoS One. 2011 Feb 18;6(2):e16765. (PMID: 21364738) ; Plant Biotechnol J. 2007 Mar;5(2):263-274. (PMID: 17309681) ; Biotechniques. 1994 Apr;16(4):664-8, 670. (PMID: 8024787) ; Nat Genet. 2005 May;37(5):501-6. (PMID: 15806101) ; Science. 2012 Nov 23;338(6110):1093-7. (PMID: 23180860) ; Plant Physiol. 1994 Oct;106(2):447-58. (PMID: 7991679) ; Plant Mol Biol. 2010 Apr;72(6):597-605. (PMID: 20076992) ; J Sci Food Agric. 2017 Oct;97(13):4273-4284. (PMID: 28182297) ; Dev Cell. 2013 May 13;25(3):317-23. (PMID: 23673333) ; Nucleic Acids Res. 1995 Feb 11;23(3):485-90. (PMID: 7885845) ; Trends Biotechnol. 2003 Dec;21(12):550-5. (PMID: 14624864) ; Plant Cell Environ. 2016 Apr;39(4):908-17. (PMID: 26670088) ; Dev Cell. 2015 Jul 27;34(2):242-51. (PMID: 26166301) ; Plant Mol Biol. 1993 Mar;21(5):895-906. (PMID: 8385509) ; Mol Gen Genet. 1990 Sep;223(3):369-78. (PMID: 2176714) ; Funct Integr Genomics. 2020 Jan;20(1):151-162. (PMID: 30796544) ; Nat Biotechnol. 2001 Feb;19(2):157-61. (PMID: 11175731) ; Bioinformatics. 2015 Oct 1;31(19):3210-2. (PMID: 26059717) ; Plant J. 2017 Mar;89(5):1042-1054. (PMID: 27775877) ; Plant Biotechnol J. 2006 Jul;4(4):445-52. (PMID: 17177809) ; Plant Mol Biol. 1994 Jan;24(1):119-27. (PMID: 8111011) ; Plant Cell. 2016 Sep;28(9):1998-2015. (PMID: 27600536) ; Plant J. 1995 Apr;7(4):649-59. (PMID: 7742860) ; Transgenic Res. 2008 Dec;17(6):1035-43. (PMID: 18437520) ; Plant Reprod. 2013 Jun;26(2):125-37. (PMID: 23539301) ; PLoS One. 2007 Aug 08;2(8):e718. (PMID: 17684564) ; Dev Cell. 2004 Jan;6(1):7-28. (PMID: 14723844) ; Plant J. 1997 May;11(5):1017-27. (PMID: 9193073) ; Plant Physiol. 2007 Dec;145(4):1220-31. (PMID: 17965180) ; Funct Plant Biol. 2003 May;30(4):443-452. (PMID: 32689029) ; Plant Biotechnol J. 2016 Feb;14(2):471-82. (PMID: 26332944) ; Plant Physiol. 2015 Oct;169(2):960-70. (PMID: 26294043) ; Development. 2001 Nov;128(21):4289-99. (PMID: 11684664) ; Plant Mol Biol. 2001 Jan;45(1):41-9. (PMID: 11247605) ; Theor Appl Genet. 2002 Mar;104(4):518-525. (PMID: 12582653) ; Annu Rev Biophys Biomol Struct. 2001;30:87-104. (PMID: 11340053) ; Nat Biotechnol. 2004 Dec;22(12):1567-72. (PMID: 15558047) ; Transgenic Res. 2007 Dec;16(6):771-81. (PMID: 17273915) ; Nucleic Acids Res. 1986 Jul 25;14(14):5641-50. (PMID: 3016659) ; Methods Mol Biol. 2006;343:143-54. (PMID: 16988341) ; Genes (Basel). 2020 Sep 12;11(9):. (PMID: 32932590) ; Plant J. 2019 Jun;98(5):767-782. (PMID: 31017340) ; In Vitro Cell Dev Biol Plant. 2016;52(4):354-366. (PMID: 27746666) ; Mol Biol Evol. 2018 Mar 1;35(3):543-548. (PMID: 29220515) ; Plant Mol Biol. 2003 Jan;51(2):263-79. (PMID: 12602884) ; Plant Cell Physiol. 2017 Jan 1;58(1):46-56. (PMID: 27856772) Grant Information: OPP1076280 bill and melinda gates foundation Contributed Indexing: Keywords: AtDD45; AtRps5a; Cowpea; Cre/lox; Promoter; Tobacco Substance Nomenclature: 0 (Plant Proteins) ; EC 2.7.7.- (Cre recombinase) ; EC 2.7.7.- (Integrases) Entry Date(s): Date Created: 20210930 Date Completed: 20220208 Latest Revision: 20220430 Update Code: 20231215 PubMed Central ID: PMC8803690

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Gene activation via Cre/lox-mediated excision in cowpea (Vigna unguiculata).