NEW METHOD OF SELECTION OF ALGAL-TRANSFORMED CELLS USING NUCLEASE

摘要

The invention relates to a method to select transformed cells. In particular, the present invention relates to the use of a nuclease engineered to inactivate selectable marker which confers cell resistance to a toxic compound. The present invention relates to methods of modifying genome of a cell, preferably an algal cell comprising the present selection step. The present invention also relates to specific engineered nucleases, polynucleotides, vectors encoding thereof, kits and isolated cells comprising said nuclease.

主权项

1. A method of modifying a algal cell comprising:
(a) Selecting a selectable marker gene within the genome of a cell which encodes a protein rendering a cell sensitive to a toxic substrate; (b) Providing a nuclease which specifically recognizes and cleaves a target sequence within said selectable marker gene; (c) Introducing said nuclease into a cell such that said nuclease cleavage inactivates said selectable marker gene; (d) Culturing said cell with said toxic substrate and; (e) Selecting cells which are resistant to the toxic substrate.