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Production method for genetically modified plant cells

A technology of transgenic plants and plant cells, applied in the fields of botanical equipment and methods, biochemical equipment and methods, plant products, etc., which can solve problems such as ambiguity and unreported possibility of mutation introduction efficiency

Inactive Publication Date: 2012-08-15
NAT INST OF AGROBIOLOGICAL SCI
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

Therefore, it is not yet clear whether zinc finger nucleases can effectively break and induce mutations at the target sites of endogenous genes in Arabidopsis.
In addition, it has recently been reported that the target sites of endogenous genes can be mutagenized by zinc finger nucleases in Zea genus and Nicotiana (Non-Patent Documents 25, 26), but there is no report on the possibility of improving the efficiency of mutation introduction

Method used

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  • Production method for genetically modified plant cells
  • Production method for genetically modified plant cells
  • Production method for genetically modified plant cells

Examples

Experimental program
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Effect test

Embodiment 1

[0099] (Example 1) Introducing a mutation into the ABI4 gene of Arabidopsis using zinc finger nuclease and the effect of the Ku80 deletion of the introduced mutation

[0100] To demonstrate zinc finger-mediated site-directed mutagenesis in Arabidopsis, the inventors chose the ABI4 gene. The present inventors found a complete consensus target site of zinc finger nuclease in ABI4 [5'-NNCNNNNNNNNNNNGNNNGNN-3' (N=A, C, G and T) / SEQ ID NO: 5].

[0101] In this example, in the construction of the modules for the construction of zinc finger nucleases, some changes were made to the method reported by Wright et al. (Wright DA, ea al. Nat Protoc 1:1637-1652, 2006). Methods. For ZF-AAA (with GGA GGA GGA as the target), the amino acid sequences near the specificity determining residues refer to 1 "QRAHLER / SEQ ID NO. 6", 2 "QSGHLQR / SEQ ID NO. 7" and 3 "QRAHLER / SEQ ID NO. 8", for ZF-TCC (with GTG GCG GCG target), it is finger 1 "RSDALTR / sequence number 9", finger 2 "RSDDLQR / serial number ...

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Abstract

By using a plant cell wherein the functioning of the protein involved in non-homologous end joining repair is artificially inhibited, the transduction-efficiency for mutations is dramatically increased in the non-homologous end joining repair process which is subsequent to the inducement of DNA double-strand breaks by zinc finger nucleases.

Description

technical field [0001] The present invention relates to a method for producing plant cells in which mutations have been introduced at specific target DNA sites on chromosomes. In more detail, it relates to: in the method of producing plant cells in which mutations have been introduced at target DNA sites on chromosomes by means of DNA double-strand breaks by restriction endonucleases and non-homologous end joining, using the A method for plant cells in which the function of double-stranded DNA repair proteins has been inhibited. Background technique [0002] The main focus of plant biotechnology is the genetic modification and improvement of crops. For this purpose, large-scale genome analysis has been accomplished in Arabidopsis as a model plant, and / or in many crops including Oryza, Zea, Triticum, Glycine, and tomato. As a result, a huge amount of genome sequence information can be utilized, and the need to develop a method capable of directly modifying a target gene in ...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N5/10A01H5/00A01H5/08A01H5/10C12N15/09
CPCA01H1/06C12N15/8213C12N15/8241C12N15/01
Inventor 土岐精一刑部敬史
Owner NAT INST OF AGROBIOLOGICAL SCI
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