Fish breeding method for improving CRISPR-Cas9 gene editing and passage efficiency through fish roe preserving fluid

A technology of gene editing and preservation solution, applied in the field of fish breeding, can solve the problems of not being able to improve the targeting efficiency and gene editing passaging efficiency of CRISPR-Cas9 gene editing technology, and achieve the goal of promoting rapid development, improving targeting efficiency and saving The effect of filter time

Active Publication Date: 2016-12-07
CHONGQING INST OF GREEN & INTELLIGENT TECH CHINESE ACADEMY OF SCI
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Problems solved by technology

[0003] However, none of the above-mentioned methods in the prior art can improve the targeting efficiency and gene editing and passage efficiency of CRISPR-Cas9 gene editing technology well.

Method used

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  • Fish breeding method for improving CRISPR-Cas9 gene editing and passage efficiency through fish roe preserving fluid
  • Fish breeding method for improving CRISPR-Cas9 gene editing and passage efficiency through fish roe preserving fluid
  • Fish breeding method for improving CRISPR-Cas9 gene editing and passage efficiency through fish roe preserving fluid

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Embodiment 1

[0034] 1. Preparation of fish roe preservation solution

[0035] Prepare fish roe preservation solution:

[0036] Take commercial Leibovitz's L-15 culture solution, add sterilized deionized water and bovine serum albumin, adjust the pH to 9.0 with 10mM NaOH, and obtain fish roe preservation solution. In the fish roe preservation solution, the volume percentage of Leibovitz's L-15 culture solution is 90%, and the final concentration of bovine serum albumin is 0.5 mg / mL.

[0037] 2. Artificial egg retrieval

[0038] Select 3-month-old male and female zebrafish with strong physique and normal development, and carry out separate tank culture in the zebrafish breeding water circulation system. The photoperiod is 14:10 (light 14h, dark 10h). Feed water worms. On the eve of the test, select female zebrafish with significantly enlarged abdomen and healthy male fish and separate them in the spawning box with a partition. The next morning, take the female fish and anesthetize them in...

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Abstract

The invention belongs to the field of fish molecular breeding, and particularly relates to a fish breeding method for improving CRISPR-Cas9 gene editing and passage efficiency through fish roe preserving fluid. According to the method, the fish roe preserving fluid technology, the microinjection technology an the CRISPR-Cas9 gene editing technology are ingeniously combined, the targeting efficiency of the CRISPR-Cas9 gene editing technology and gene editing passage efficiency are greatly improved, the screening time needed by a gene editing method for fish breeding is obviously saved, and the method is of great significance in promoting rapid development of gene editing fish breeding.

Description

technical field [0001] The invention belongs to the field of fish molecular breeding, and in particular relates to a fish breeding method for improving CRISPR-Cas9 gene editing and passage efficiency by using a fish egg preservation solution. Background technique [0002] Gene editing is of great significance to the study of fish gene functions and fish breeding and species improvement. However, the traditional CRISPR-Cas9 gene editing method has low targeting efficiency, which brings heavy work to the screening of gene-edited fish and seriously hinders the development of gene-edited fish breeding. In recent years, many scholars have devoted themselves to the research of methods to improve the efficiency of gene editing and passage efficiency, such as co-injecting fluorescent proteins that can be specifically expressed in gonads during microinjection to achieve fluorescent labeling of germ cells (Dong Z, Dong X, Jia W, et al., The international journal of biochemistry & cel...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N15/89A01K67/027
CPCA01K67/0276A01K2227/40C12N15/89
Inventor 裴得胜谢少林
Owner CHONGQING INST OF GREEN & INTELLIGENT TECH CHINESE ACADEMY OF SCI
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