Plant-derived cytosine deaminases and application thereof to base editing system

A technology of cytosine deaminase and base editing, applied in the application field of gene editing, can solve the application of cytosine deaminase that has not been studied, the application of cytosine deaminase that has not been studied, the difference in function and tissue-specific expression And other issues

Active Publication Date: 2021-01-19
CROPEDIT BIOTECHNOLOGY INC
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

There are many studies on CD in plants, but there has never been any research on the application of CD from plants in gene editing technology, and there is no CD from plants at home and abroad. Reports on the application of gene editing technology
In the study of the cytosine deaminase (APOBEC

Method used

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Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0047] Example 1: Base gene editing in rice using cytosine deaminase from Arabidopsis thaliana.

[0048] 1.1. Construction of gene editing vectors

[0049] The inventors of the present invention constructed the vector nCas9&AtCDA1&UGI. Wherein AtCDA1 is a cytosine deaminase derived from Arabidopsis thaliana, and its amino acid sequence is shown in sequence 1 in the sequence list.

[0050] The vector nCas9&AtCDA1&UGI used in this example is a circular plasmid, and its nucleotide sequence is shown as sequence 47 in the sequence listing. In the above sequence, the 93rd to 277th position is the nucleotide sequence of the PolyA terminator, the 318th to 1343rd position is the nucleotide sequence of the hygromycin phosphotransferase, and the 1378th to 2157th position is the nucleotide sequence of the 35S promoter Sequence, the 2452nd to 2706th is the nucleotide sequence of NOS terminator, the 2743rd to 2991st is the nucleotide sequence of UGI, the 3061st to 7161st is the nucleotid...

Embodiment 2

[0068] Example 2: Base gene editing in rice using cytosine deaminase APOBEC1 derived from rice.

[0069] 2.1 Construction of gene editing vector

[0070] The inventors of the present invention constructed the vector nCas9&APOBEC1&UGI. Wherein APOBEC1 is a cytosine deaminase derived from rice, and its amino acid sequence is shown in sequence 2 in the sequence list.

[0071] The vector nCas9&APOBEC1&UGI used in this example is a circular plasmid, and its nucleotide sequence is shown in sequence 48 in the sequence listing. Among the above sequences, the 93rd to 277th are the nucleotide sequence of the PolyA terminator, the 318th to 1343rd are the nucleotide sequence of the hygromycin, the 1378th to 2157th are the nucleotide sequence of the 35S promoter, and the 35S promoter is the nucleotide sequence of the 35S promoter. 2452 to 2706 are the nucleotide sequence of the NOS terminator, the 2743 to 2991 are the nucleotide sequence of UGI, the 3061 to 7161 are the nucleotide sequen...

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PUM

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Abstract

The invention discloses plant-derived cytosine deaminases and a method and application thereof to a base editing system. Experiments prove that the plant-derived cytosine deaminase can successfully perform base editing on receptor genes, and has important value for application of gene editing to plants.

Description

technical field [0001] The invention belongs to the technical field of application of genes, and specifically relates to the gene editing of plants in the field of biotechnology, using plant-derived cytosine deaminase instead of non-plant-derived cytosine deaminase in a base editing system Applications. Background technique [0002] As one of the rapidly developing biotechnologies in recent years, gene editing technology has been widely used in animals, plants and microorganisms, and the exploration of the principles of this technology or the expansion of its applications are also being actively carried out. Gene editing technology is to replace, excise, add or insert exogenous DNA sequences to a nucleotide sequence or even a single nucleotide sequence of the target gene in the cell genome to produce heritable changes. [1]- [3] Although the principles and modes of action of various gene editing technologies are different, what they have in common is that gene editing is b...

Claims

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Application Information

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IPC IPC(8): C12N9/78C12N15/55C12N15/82A01H5/00A01H6/46
CPCC12N9/78C12Y305/04001C12N15/8213
Inventor 张蕾李相敢侯丽敏祁幼林王身昌黎跃进
Owner CROPEDIT BIOTECHNOLOGY INC
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