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Application of SpRYn-CBE base editing system in plant genome base replacement

A plant genome and coding gene technology, applied in the application field of SpRYn-CBE base editing system in plant genome base replacement, can solve problems such as limited range

Pending Publication Date: 2022-04-12
BEIJING ACADEMY OF AGRICULTURE & FORESTRY SCIENCES
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0004] At present, the SpCas9n(D10A)&rAPOBEC1 / PmCDA1&UGI base editing system has been widely used in rice to realize the conversion of C to T, but the editing target is mainly limited to the sequence of PAM (Protospacer Adjacent Motif) as NGG, which greatly limits the Editable range of C

Method used

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  • Application of SpRYn-CBE base editing system in plant genome base replacement
  • Application of SpRYn-CBE base editing system in plant genome base replacement
  • Application of SpRYn-CBE base editing system in plant genome base replacement

Examples

Experimental program
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Embodiment 1

[0147] Example 1. Construction of the SpRYn-CBE base editing system vector and its application in the base substitution of targets with PAM sequences of NAN, NCN or NTN in the rice genome

[0148] 1. Construction of SpRYn-CBE base editing system vector

[0149] Artificially synthesize the following recombinant expression vectors, each of which is a circular plasmid: SpRYn-CBE-1 recombinant expression vector, SpRYn-CBE-2 recombinant expression vector, SpRYn-CBE-3 recombinant expression vector, SpRYn-CBE-4 recombinant expression vector , SpRYn-CBE-5 recombinant expression vector, SpRYn-CBE-6 recombinant expression vector, SpRYn-CBE-7 recombinant expression vector, SpRYn-CBE-8 recombinant expression vector, SpRYn-CBE-9 recombinant expression vector, SpRYn-CBE- 10 recombinant expression vectors, SpRYn-CBE-11 recombinant expression vector, SpRYn-CBE-12 recombinant expression vector, SpRYn-CBE-13 recombinant expression vector, SpRYn-CBE-14 recombinant expression vector, SpRYn-CBE-15...

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Abstract

The invention discloses an application of a SpRYn-CBE base editing system in plant genome base replacement. The SpRYn-CBE basic group editing system disclosed by the invention comprises SpRYn, PmCDA1, UGI and sgRNA (Single Guide Ribonucleic Acid), the sgRNA is targeted to a target spot sequence. Experiments prove that the SpRYn-CBE base editing system disclosed by the invention can be used for editing a base C in a target sequence of which the PAM sequence is NAN or NCN or NTN on a plant genome, so that the base C is replaced into a base T, and the range of editable C is greatly expanded.

Description

technical field [0001] The invention relates to the field of biotechnology, in particular to the application of the SpRYn-CBE base editing system in plant genome base replacement. Background technique [0002] CRISPR-Cas9 technology has become a powerful genome editing method and has been widely used in many tissues and cells. The CRISPR / Cas9 protein-RNA complex is positioned on the target by the guide RNA (guide RNA), cuts and generates a DNA double-strand break (dsDNA break, DSB), and then the organism will instinctively initiate a DNA repair mechanism to repair the DSB. There are generally two repair mechanisms, one is non-homologous end joining (NHEJ), and the other is homologous recombination (homology-directed repair, HDR). Usually NHEJ accounts for the majority, so the random indels (insertions or deletions) generated by the repair are much higher than the precise repair. For precise base substitution, the application of HDR to achieve precise base substitution is g...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N15/10C12N15/82C12N15/55C12N9/78A01H5/00A01H6/46
Inventor 杨进孝赵久然王飞鹏王瑶张成伟
Owner BEIJING ACADEMY OF AGRICULTURE & FORESTRY SCIENCES