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Main Authors: Yan, Junjun, Qian, Siyu, Li, Guo, Liu, Yujie, Zhou, Liqian, Ye, Tiantian, Liu, Cui, Meng, Jilun, Jie, Yukun, Gu, Zhimin
Format: Artículo científico
Language:en
Published: Animals : an open access journal from MDPI 2025
Online Access:https://pubmed.ncbi.nlm.nih.gov/41514701/
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author Yan, Junjun
Qian, Siyu
Li, Guo
Liu, Yujie
Zhou, Liqian
Ye, Tiantian
Liu, Cui
Meng, Jilun
Jie, Yukun
Gu, Zhimin
author_facet Yan, Junjun
Qian, Siyu
Li, Guo
Liu, Yujie
Zhou, Liqian
Ye, Tiantian
Liu, Cui
Meng, Jilun
Jie, Yukun
Gu, Zhimin
Yan, Junjun
Qian, Siyu
Li, Guo
Liu, Yujie
Zhou, Liqian
Ye, Tiantian
Liu, Cui
Meng, Jilun
Jie, Yukun
Gu, Zhimin
collection PubMed - marine biology
contents Establishment of a CRISPR/Cas9-Based Genome Editing System in . Yan, Junjun Qian, Siyu Li, Guo Liu, Yujie Zhou, Liqian Ye, Tiantian Liu, Cui Meng, Jilun Jie, Yukun Gu, Zhimin The giant freshwater prawn is a globally farmed decapod species of high economic and nutritional value, but its genetic improvement has been constrained by the lack of an efficient genome editing workflow. Here, we optimized a CRISPR/Cas9 microinjection system for targeted mutagenesis in embryos. A controlled mating-based approach enabled routine collection of one-cell embryos, and developmental staging identified 0.5-2 h post-fertilization as the optimal injection window. Optimization of the needle tip (~1 μm) minimized mechanical damage, lowering early embryo mortality to ~10%. Using this system, the eye-development gene and the sex-related gene were successfully edited. editing produced a 46.9% mutation rate and an average 50% reduction in eye pigment area. editing achieved an 84% mutation rate, while post-injection survival to hatching was approximately 35%. These results establish an efficient and reproducible CRISPR/Cas9 editing system in , providing a foundation for functional genomics and genetic improvement in this commercially important prawn species.
format Artículo científico
id pubmed_41514701
institution PubMed
language en
publishDate 2025
publisher Animals : an open access journal from MDPI
record_format pubmed
spellingShingle Establishment of a CRISPR/Cas9-Based Genome Editing System in .
Yan, Junjun
Qian, Siyu
Li, Guo
Liu, Yujie
Zhou, Liqian
Ye, Tiantian
Liu, Cui
Meng, Jilun
Jie, Yukun
Gu, Zhimin
Establishment of a CRISPR/Cas9-Based Genome Editing System in . Yan, Junjun Qian, Siyu Li, Guo Liu, Yujie Zhou, Liqian Ye, Tiantian Liu, Cui Meng, Jilun Jie, Yukun Gu, Zhimin The giant freshwater prawn is a globally farmed decapod species of high economic and nutritional value, but its genetic improvement has been constrained by the lack of an efficient genome editing workflow. Here, we optimized a CRISPR/Cas9 microinjection system for targeted mutagenesis in embryos. A controlled mating-based approach enabled routine collection of one-cell embryos, and developmental staging identified 0.5-2 h post-fertilization as the optimal injection window. Optimization of the needle tip (~1 μm) minimized mechanical damage, lowering early embryo mortality to ~10%. Using this system, the eye-development gene and the sex-related gene were successfully edited. editing produced a 46.9% mutation rate and an average 50% reduction in eye pigment area. editing achieved an 84% mutation rate, while post-injection survival to hatching was approximately 35%. These results establish an efficient and reproducible CRISPR/Cas9 editing system in , providing a foundation for functional genomics and genetic improvement in this commercially important prawn species.
title Establishment of a CRISPR/Cas9-Based Genome Editing System in .
url https://pubmed.ncbi.nlm.nih.gov/41514701/