Split-Cre complementation restores combination activity on transgene excision in hair roots of transgenic tobacco.

The Cre/loxP system is increasingly exploited for genetic manipulation of DNA in vitro and in vivo. It was previously reported that inactive ''split-Cre'' fragments could restore Cre activity in transgenic mice when overlapping co-expression was controlled by two different promot...

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Autores principales:	Mengling Wen, Yuan Gao, Lijun Wang, Lingyu Ran, Jiahui Li, Keming Luo
Formato:	article
Lenguaje:	EN
Publicado:	Public Library of Science (PLoS) 2014
Materias:	Medicine R Science Q
Acceso en línea:	https://doaj.org/article/08565637f9a34f6ab4e416cc01cb6f43
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spelling	oai:doaj.org-article:08565637f9a34f6ab4e416cc01cb6f432021-11-25T05:56:11ZSplit-Cre complementation restores combination activity on transgene excision in hair roots of transgenic tobacco.1932-620310.1371/journal.pone.0110290https://doaj.org/article/08565637f9a34f6ab4e416cc01cb6f432014-01-01T00:00:00Zhttps://doi.org/10.1371/journal.pone.0110290https://doaj.org/toc/1932-6203The Cre/loxP system is increasingly exploited for genetic manipulation of DNA in vitro and in vivo. It was previously reported that inactive ''split-Cre'' fragments could restore Cre activity in transgenic mice when overlapping co-expression was controlled by two different promoters. In this study, we analyzed recombination activities of split-Cre proteins, and found that no recombinase activity was detected in the in vitro recombination reaction in which only the N-terminal domain (NCre) of split-Cre protein was expressed, whereas recombination activity was obtained when the C-terminal (CCre) or both NCre and CCre fragments were supplied. We have also determined the recombination efficiency of split-Cre proteins which were co-expressed in hair roots of transgenic tobacco. No Cre recombination event was observed in hair roots of transgenic tobacco when the NCre or CCre genes were expressed alone. In contrast, an efficient recombination event was found in transgenic hairy roots co-expressing both inactive split-Cre genes. Moreover, the restored recombination efficiency of split-Cre proteins fused with the nuclear localization sequence (NLS) was higher than that of intact Cre in transgenic lines. Thus, DNA recombination mediated by split-Cre proteins provides an alternative method for spatial and temporal regulation of gene expression in transgenic plants.Mengling WenYuan GaoLijun WangLingyu RanJiahui LiKeming LuoPublic Library of Science (PLoS)articleMedicineRScienceQENPLoS ONE, Vol 9, Iss 10, p e110290 (2014)
institution	DOAJ
collection	DOAJ
language	EN
topic	Medicine R Science Q
spellingShingle	Medicine R Science Q Mengling Wen Yuan Gao Lijun Wang Lingyu Ran Jiahui Li Keming Luo Split-Cre complementation restores combination activity on transgene excision in hair roots of transgenic tobacco.
description	The Cre/loxP system is increasingly exploited for genetic manipulation of DNA in vitro and in vivo. It was previously reported that inactive ''split-Cre'' fragments could restore Cre activity in transgenic mice when overlapping co-expression was controlled by two different promoters. In this study, we analyzed recombination activities of split-Cre proteins, and found that no recombinase activity was detected in the in vitro recombination reaction in which only the N-terminal domain (NCre) of split-Cre protein was expressed, whereas recombination activity was obtained when the C-terminal (CCre) or both NCre and CCre fragments were supplied. We have also determined the recombination efficiency of split-Cre proteins which were co-expressed in hair roots of transgenic tobacco. No Cre recombination event was observed in hair roots of transgenic tobacco when the NCre or CCre genes were expressed alone. In contrast, an efficient recombination event was found in transgenic hairy roots co-expressing both inactive split-Cre genes. Moreover, the restored recombination efficiency of split-Cre proteins fused with the nuclear localization sequence (NLS) was higher than that of intact Cre in transgenic lines. Thus, DNA recombination mediated by split-Cre proteins provides an alternative method for spatial and temporal regulation of gene expression in transgenic plants.
format	article
author	Mengling Wen Yuan Gao Lijun Wang Lingyu Ran Jiahui Li Keming Luo
author_facet	Mengling Wen Yuan Gao Lijun Wang Lingyu Ran Jiahui Li Keming Luo
author_sort	Mengling Wen
title	Split-Cre complementation restores combination activity on transgene excision in hair roots of transgenic tobacco.
title_short	Split-Cre complementation restores combination activity on transgene excision in hair roots of transgenic tobacco.
title_full	Split-Cre complementation restores combination activity on transgene excision in hair roots of transgenic tobacco.
title_fullStr	Split-Cre complementation restores combination activity on transgene excision in hair roots of transgenic tobacco.
title_full_unstemmed	Split-Cre complementation restores combination activity on transgene excision in hair roots of transgenic tobacco.
title_sort	split-cre complementation restores combination activity on transgene excision in hair roots of transgenic tobacco.
publisher	Public Library of Science (PLoS)
publishDate	2014
url	https://doaj.org/article/08565637f9a34f6ab4e416cc01cb6f43
work_keys_str_mv	AT menglingwen splitcrecomplementationrestorescombinationactivityontransgeneexcisioninhairrootsoftransgenictobacco AT yuangao splitcrecomplementationrestorescombinationactivityontransgeneexcisioninhairrootsoftransgenictobacco AT lijunwang splitcrecomplementationrestorescombinationactivityontransgeneexcisioninhairrootsoftransgenictobacco AT lingyuran splitcrecomplementationrestorescombinationactivityontransgeneexcisioninhairrootsoftransgenictobacco AT jiahuili splitcrecomplementationrestorescombinationactivityontransgeneexcisioninhairrootsoftransgenictobacco AT kemingluo splitcrecomplementationrestorescombinationactivityontransgeneexcisioninhairrootsoftransgenictobacco
_version_	1718414322028249088

Split-Cre complementation restores combination activity on transgene excision in hair roots of transgenic tobacco.

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