Polypeptides and nucleic acids that mediate endocytosis of exogenous proteins and their applications and methods for gene editing in Neopodia prawns

A foreign protein and nucleic acid technology, applied in the field of gene editing, can solve problems such as difficult expression, purification, no practical application value, and large molecular weight of fusion proteins

Active Publication Date: 2022-04-22
TIANJIN NORMAL UNIVERSITY
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

However, the molecular weight of crustacean YPPs is as high as 300kDa, and the fusion protein formed by fusing it as a YPP ligand with Cas9 is too large in molecular weight, making it difficult to express, purify, form RNP complexes and be endocytosed, so it has no practical application value.

Method used

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  • Polypeptides and nucleic acids that mediate endocytosis of exogenous proteins and their applications and methods for gene editing in Neopodia prawns
  • Polypeptides and nucleic acids that mediate endocytosis of exogenous proteins and their applications and methods for gene editing in Neopodia prawns
  • Polypeptides and nucleic acids that mediate endocytosis of exogenous proteins and their applications and methods for gene editing in Neopodia prawns

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Embodiment Construction

[0027] In order to enable those skilled in the art to better understand the solution of the present invention, the technical solution of the present invention will be further described below in conjunction with specific examples.

[0028] The experimental methods in the following examples are conventional methods unless otherwise specified. The test materials used in the following examples, unless otherwise specified, were purchased from conventional biochemical reagent stores. % in the following examples, unless otherwise specified, are mass percentages. The biological material used in the embodiment is the new rice shrimp purchased from Zhejiang Aquatic Market, with a body length of about 2 cm, healthy and vigorous. The rice shrimp was raised in the laboratory after being bought back. Fresh water aeration, the temperature is about 26-28 ℃. Shrimp food was fed once a day, and fresh aeration water was replaced every two weeks.

[0029] Step 1. Bioinformatics analysis of vi...

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Abstract

The invention discloses polypeptides and nucleic acids that mediate the endocytosis of exogenous proteins and their application, and a method for gene editing in Neocaridina heteropoda. The invention finds a length of A polypeptide of only 40aa, the polypeptide has an unexpected effect of mediating the endocytosis of exogenous proteins, so it is easy to express, purify, form RNP complexes and be endocytosed, and has a good effect in the gene editing of Neopodia prawn Application prospects. This technology can be widely applied to non-model organisms that are not yet proficient in embryonic microinjection technology, and is of great significance for Cas9-mediated gene editing.

Description

technical field [0001] The invention belongs to the field of biotechnology, and specifically relates to polypeptides and nucleic acids that mediate endocytosis of exogenous proteins and their applications, and a method for gene editing in Neopodia prawns. Background technique [0002] CRISPR / Cas9 (Clustered regulatory interspaced short palindromicrepeat / Cas9) system is currently the most popular method in the field of genome editing. CRISPR is a special DNA repeat sequence family widely distributed in the genomes of bacteria and archaea. The restriction site of CRISPR is usually composed of highly conserved repeat sequences with a length of 21-48bp and 26-72bp between repeat sequences. These spacers are the markers for CRISPR to recognize the target gene. Cas9 exists near the CRISPR site and is a double-stranded DNA nuclease that can recognize and cut near the PAM (protospacer adjacent motif) sequence adjacent to the target site under the guidance of sgRNA (small guide RNA)...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C07K14/435C12N15/12C12N15/90C12N9/22A01K67/033
CPCC07K14/43509C12N15/902C12N9/22A01K67/0338A01K2207/10A01K2217/07A01K2227/70
Inventor 李冉王鑫孙金生
Owner TIANJIN NORMAL UNIVERSITY
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