'GM-gene-deletor': fused loxP-FRT recognition sequences dramatically improve the efficiency of FLP or CRE recombinase on transgene excision from pollen and seed of tobacco plants

doi:10.1111/j.1467-7652.2006.00237.x

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Wiley InterScience

Plant Biotechnology Journal

Volume 5 Issue 2, Pages 263 - 374

Published Online: 26 Jan 2007

Plant Biotechnology Journal is published by Wiley-Blackwell in association with the Society for Experimental Biology (SEB) and the Association of Applied Biologists (AAB).

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'GM-gene-deletor': fused loxP-FRT recognition sequences dramatically improve the efficiency of FLP or CRE recombinase on transgene excision from pollen and seed of tobacco plants

Keming Luo ^1,2,†, Hui Duan ^1,†^,‡, Degang Zhao ^1,†^,§, Xuelian Zheng ¹ , Wei Deng ¹ , Yongqin Chen ^1,¶, C. Neal Stewart Jr ³ , Richard McAvoy ¹ , Xiangning Jiang ^1,**, Yanhong Wu ¹ , Aigong He ^1,§, Yan Pei ^2,* and Yi Li ^1,*

¹ Department of Plant Science, University of Connecticut, Storrs, CT 06269, USA
² Biotechnology Center, Southwest University, Chongqing 400716, China
³ Department of Plant Sciences, University of Tennessee, Knoxville, TN 37996-4561, USA

* Correspondence (fax +1 860 486 6777; e-mail yi.li@uconn.edu) or (fax +1 86 23 63250515; e-mail peiyan@swu.edu.cn)

†These authors contributed equally to this work

‡ Present address: Department of Cell and Structural Biology, University of Illinois, Urbana, IL 61801, USA

§Visiting scientists (postdoctorates) from Guizhou University, Guizhou, China

¶Visiting scientist from Hubei University, Hubei, China

**Visiting scientist from Beijing Forestry University, Beijing, China

KEYWORDS

biosafety • CRE/loxP • FLP/ FRT • loxP-FRT fusion recognition site • non-transgenic pollen and seed • transgene flow

ABSTRACT

Pollen- and seed-mediated transgene flow is a concern in plant biotechnology. We report here a highly efficient 'genetically modified (GM)-gene-deletor' system to remove all functional transgenes from pollen, seed or both. With the three pollen- and/or seed-specific gene promoters tested, the phage CRE/loxP or yeast FLP/FRT system alone was inefficient in excising transgenes from tobacco pollen and/or seed, with no transgenic event having 100% efficiency. When loxP-FRT fusion sequences were used as recognition sites, simultaneous expression of both FLP and CRE reduced the average excision efficiency, but the expression of FLP or CRE alone increased the average excision efficiency, with many transgenic events being 100% efficient based on more than 25 000 T₁ progeny examined per event. The 'GM-gene-deletor' reported here may be used to produce 'non-transgenic' pollen and/or seed from transgenic plants and to provide a bioconfinement tool for transgenic crops and perennials, with special applicability towards vegetatively propagated plants and trees.

Received 22 August 2006; revised 20 October 2006; accepted 26 October 2006.

DIGITAL OBJECT IDENTIFIER (DOI)

10.1111/j.1467-7652.2006.00237.x About DOI