Rice uracil DNA glycosidase and application thereof in gene editing induction of plant single base diversity

A technology of uracil and glycosidase, which is applied in the fields of biotechnology and plant genetic engineering, can solve the problems of limited base types, low efficiency of fixed-point replacement, and limit the scope of application of single base editors, so as to expand the types of mutations and crops Genetic diversity and the effect of expanding the scope of application

Pending Publication Date: 2022-08-05
CAS CENT FOR EXCELLENCE IN MOLECULAR PLANT SCI
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

At present, the traditional plant gene cytosine single base editing system mainly induces the single-base substitution of cytosine C->thymine T, and because cytosine C->guanine G or cytosine C->adenine A fixed-point replacement efficiency is very low, and the types of bases that can be replaced are relatively limited, which greatly limits the scope of application of single base editors

Method used

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  • Rice uracil DNA glycosidase and application thereof in gene editing induction of plant single base diversity
  • Rice uracil DNA glycosidase and application thereof in gene editing induction of plant single base diversity
  • Rice uracil DNA glycosidase and application thereof in gene editing induction of plant single base diversity

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Experimental program
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Embodiment 1

[0037] see attached Figure 1-8 , the rice uracil DNA glycosidase OsUNG provided in the embodiment of the present invention is a protein composed of the amino acid sequence shown in SEQ ID NO.2; or the amino acid sequence of SEQ ID NO.2 is processed by a or substitution and / or deletion of several amino acid residues and / or addition of a protein derived from SEQ ID NO. 2 and retaining the function of the protein shown in SEQ ID NO. 2.

[0038] The encoding gene of the rice uracil DNA glycosidase UNG has:

[0039] A. The nucleotide sequence shown in SEQ ID No. 1, or

[0040] B. A nucleotide sequence derived from A with the equivalent function of one or more nucleotides substituted, deleted or added to the nucleotide sequence shown in SEQ ID No. 1; or

[0041] C. The nucleotide sequence that hybridizes to the sequence shown in SEQ ID NO: 1 and expresses the same functional protein under the setting of 0.1X SSPE with 0.1% SDS or 0.1X SSC with 0.1% SDS solution, hybridize at 65°...

Embodiment 2

[0074] The rice uracil DNA glycosidase and its application in gene editing to induce plant monobasic diversity provided in the embodiments of the present invention are basically the same as those in Example 1, except that:

[0075] The amino acid sequence of the rice uracil DNA glycosidase is: a protein composed of the amino acid sequence shown in SEQ ID NO.2; or the amino acid sequence of SEQ ID NO.2 is replaced by one or several amino acid residues and / or deletion and / or addition of a protein derived from SEQ ID NO. 2 that retains the function of the protein shown in SEQ ID NO. 2.

[0076] The encoding gene of the rice uracil DNA glycosidase UNG has:

[0077] E. The nucleotide sequence shown in SEQ ID No. 1, or

[0078] F. The nucleotide sequence derived from E with the equivalent function in the nucleotide sequence shown in SEQ ID No. 1 by substitution, deletion or addition of one or more nucleotides; or

[0079] G. The nucleotide sequence that hybridizes to the sequence...

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Abstract

The invention discloses rice uracil DNA (deoxyribonucleic acid) glycosidase and application thereof in gene editing induction of plant single base diversity. The rice uracil DNA glycosidase is a protein composed of an amino acid sequence as shown in SEQ ID NO. 2; or a protein which is derived from the amino acid sequence shown in SEQ ID NO.2 and keeps the protein function shown in SEQ ID NO.2 through substitution and / or deletion and / or addition of one or more amino acid residues of the amino acid sequence shown in SEQ ID NO.2. According to the application provided by the invention, a biological material containing the rice uracil DNA glycosidase UNG is constructed into a plant cytosine single-base editing vector OsCGBE containing the OsUNG sequence, the expression vector is introduced into a plant, a C base on DNA is induced to be mutated into T or G or A, plant single-base diversity is efficiently created, and the application range of a single-base editor is expanded.

Description

technical field [0001] The invention relates to the technical fields of biotechnology and plant genetic engineering, in particular to a rice uracil DNA glycosidase and its application in gene editing to induce plant single base diversity. Background technique [0002] Artificial nucleases developed in recent years can effectively edit target fragments by causing DNA double-strand breaks at specific sites. Base editors, currently including cytidine base editor (CBE) and adenine base editor (ABE), enable precise base changes in the genome without inducing DNA duplication Strand breaks (DSBs). Many agronomic traits in plants are controlled by changes in one or a few DNA bases, so base editors are important tools in plant molecular breeding. [0003] Existing single-base editor CBE and ABE systems, activated by the rat cytidine deaminase APOBEC1 (rAPOBEC1), the activation-induced cytidine deaminase (AID) PmCDA1, or the evolved tRNA adenine deaminase TadA, It is fused with Cas...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N9/24C12N15/56C12N15/82C12N15/84A01H5/00A01H6/46
CPCC12N9/2497C12Y302/02027C12N15/8241C12N15/8216C12N15/8205
Inventor 朱健康陆钰明田益夫沈润东
Owner CAS CENT FOR EXCELLENCE IN MOLECULAR PLANT SCI
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