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GRNA and use thereof

A nucleic acid and repetitive sequence technology, applied in the direction of DNA / RNA fragments, recombinant DNA technology, the use of vectors to introduce foreign genetic material, etc., can solve problems such as reducing off-target effects

Active Publication Date: 2021-09-03
SHANDONG SHUNFENG BIOTECH CO LTD
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

In addition, the PAM sequences of Cas9, Cpf1, CasX, and CasY are relatively complex and diverse, while C2c1 recognizes a strict 5'-TTN, so its target site is easier to predict than other systems, thereby reducing potential off-target effects

Method used

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  • GRNA and use thereof
  • GRNA and use thereof
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Examples

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Embodiment Construction

[0047] Guide RNA (guide RNA, gRNA)

[0048] In one aspect, the present invention provides a gRNA, which includes a first segment and a second segment; the first segment is also called "skeleton region", "protein binding segment", "protein binding sequence ", or "direct repeat (DirectRepeat) sequence"; the second segment is also known as "targeting nucleic acid-targeting sequence" or "targeting nucleic acid-targeting segment".

[0049]The nucleic acid-targeting sequence or nucleic acid-targeting segment of the present invention comprises a nucleotide sequence that is complementary to a sequence in the target nucleic acid. In other words, the nucleic acid-targeting targeting sequence or nucleic acid-targeting segment of the present invention interacts with the target nucleic acid in a sequence-specific manner through hybridization (ie, base pairing). Accordingly, the nucleic acid-targeting sequence or nucleic acid-targeting segment can be altered, or modified, to hybridize to a...

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Abstract

The invention belongs to the field of nucleic acid editing and particularly relates to the technical field of clustered regularly interspaced short palindromic repeat (CRISPR). Specifically, the invention provides a gRNA and the gRNA can be used for improving efficiency of a Cas enzyme.

Description

technical field [0001] The invention relates to the field of nucleic acid editing, in particular to the technical field of clustered regularly interspaced short palindromic repeats (CRISPR). Specifically, the present invention relates to a gRNA and its use, especially to a gRNA suitable for the Cas12 protein family and its use. Background technique [0002] CRISPR / Cas technology is a widely used gene editing technology. It uses RNA guidance to specifically bind to target sequences on the genome and cuts DNA to generate double-strand breaks. It uses biological non-homologous end joining or homologous recombination for site-directed gene editing. [0003] The CRISPR / Cas9 system is the most commonly used type II CRISPR system, which recognizes the PAM motif of 3’-NGG and performs blunt-end cleavage of the target sequence. The CRISPR / Cas Type V system is a newly discovered CRISPR system in the past two years. It has a 5'-TTN motif and cuts the sticky end of the target sequence...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N15/113C12N9/22C12N15/63C12N15/82C12Q1/68
CPCC12N15/113C12N9/22C12N15/63C12N15/8218C12Q1/68C12N2310/20C12Q2521/327C12Q2525/161
Inventor 段志强梁亚峰
Owner SHANDONG SHUNFENG BIOTECH CO LTD
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