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פותח על ידי קלירמאש פתרונות בע"מ -
Genome Editing Using the CRISPR-Cas9 System to Generate a Solid-Red Germline of Nile Tilapia ( Oreochromis niloticus)
Year:
2021
Source of publication :
The CRISPR Journal
Authors :
בירן, יעקב
;
.
כנעני, אבנר
;
.
סלוסמן, טטיאנה
;
.
רוזן, עדה
;
.
שרמן, עמיר
;
.
Volume :
4
Co-Authors:

Adi Segev-Hadar 
Tatiana Slosman
Ada Rozen 
Amir Sherman
Avner Cnaani
Jakob Biran 

Facilitators :
From page:
583
To page:
594
(
Total pages:
12
)
Abstract:

In recent years, there has been increasing demand for red tilapia, which are commercial strains of hybrids of different tilapiine species or red variants of highly inbred Nile tilapia. However, red tilapia phenotypes are genetically unstable and affected by environmental factors, resulting in nonuniform coloration with black or dark-red color blotches that reduce their market value. Solute carrier family 45 member 2 (SLC45A2) is a membrane transporter that mediates melanin biosynthesis and is evolutionarily conserved from fish to humans. In the present study, we describe the generation of a stable and heritable red tilapia phenotype by inducing loss-of-function mutations in the slc45a2 gene. For this purpose, we identified the slc45a2 gene in Nile tilapia and designed highly specific guide RNAs (gRNA) for its genomic sequence. Multiplex microinjection of slc45a2-specific ribonucleoproteins to Nile tilapia zygotes induced up to 97-99% albinism, including loss of melanin in the eye. Next-generation sequencing of the injected zygotes demonstrated that all injected fish carried mutant alleles with variable mutagenesis efficiencies. Sanger sequencing of the genomic target region in the slc45a2 gene from fin clips, sperm, and F1 offspring of a highly mutant male identified various genomic indels and germline transmission of the sperm-identified indels. Overall, this work demonstrates the generation of somatic and germline slc45a2 mutant alleles, which leads to complete albinism in Nile tilapia.

Note:
Related Files :
CRISPR/Cas9
genome editing
nile tilapia (Oreochromis nilocticus)
עוד תגיות
תוכן קשור
More details
DOI :
10.1089/crispr.2020.0115
Article number:
0
Affiliations:
Database:
PubMed
Publication Type:
מאמר
;
.
Language:
אנגלית
Editors' remarks:
ID:
56071
Last updated date:
20/03/2022 10:52
Creation date:
25/08/2021 17:00
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Scientific Publication
Genome Editing Using the CRISPR-Cas9 System to Generate a Solid-Red Germline of Nile Tilapia ( Oreochromis niloticus)
4

Adi Segev-Hadar 
Tatiana Slosman
Ada Rozen 
Amir Sherman
Avner Cnaani
Jakob Biran 

Genome Editing Using the CRISPR-Cas9 System to Generate a Solid-Red Germline of Nile Tilapia ( Oreochromis niloticus)

In recent years, there has been increasing demand for red tilapia, which are commercial strains of hybrids of different tilapiine species or red variants of highly inbred Nile tilapia. However, red tilapia phenotypes are genetically unstable and affected by environmental factors, resulting in nonuniform coloration with black or dark-red color blotches that reduce their market value. Solute carrier family 45 member 2 (SLC45A2) is a membrane transporter that mediates melanin biosynthesis and is evolutionarily conserved from fish to humans. In the present study, we describe the generation of a stable and heritable red tilapia phenotype by inducing loss-of-function mutations in the slc45a2 gene. For this purpose, we identified the slc45a2 gene in Nile tilapia and designed highly specific guide RNAs (gRNA) for its genomic sequence. Multiplex microinjection of slc45a2-specific ribonucleoproteins to Nile tilapia zygotes induced up to 97-99% albinism, including loss of melanin in the eye. Next-generation sequencing of the injected zygotes demonstrated that all injected fish carried mutant alleles with variable mutagenesis efficiencies. Sanger sequencing of the genomic target region in the slc45a2 gene from fin clips, sperm, and F1 offspring of a highly mutant male identified various genomic indels and germline transmission of the sperm-identified indels. Overall, this work demonstrates the generation of somatic and germline slc45a2 mutant alleles, which leads to complete albinism in Nile tilapia.

Scientific Publication
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