Utilization of Founder Lines for Improved Citrus Biotechnology via RMCE
Subtropical Insects and Horticulture Research
Project Number: 6034-21000-014-22
Start Date: Oct 01, 2013
End Date: Sep 30, 2015
The research objectives are to reduce potentially negative effects of transgene insertion and genomic presence. The aim of this proposed research is to investigate the use of novel unidirectional recombinases Bxbl, CinH. ParA and phiC31 to implement a Recombinase-Mediate Cassette Exchange (RMCE) technique for precise integration with simultaneous marker removal.
1. To test the newly generated transgenic founder citrus Carrizo citrange lines containing the RMCE genetic platform for precise targeting.
2. To demonstrate that the dual unidirectional RMCE can be exchanged for constructs with desirable transgenes in Carrizo citrange.
Transgenic Carrizo citrange founder lines have been generated containing the
selection gene cassette flanked by fused recognition sites. Single copy-number, highly expressing lines will be selected. An exchange vector will be
transformed into the various Carrizo citrange founder lines. Recombinase
mediated cassette exchange will be examined by negative selection and used to score the most effective pairs. The most effective Carrizo citrange founder lines and recombinase pairs will be published and made publicly available.