Generation of Myostatin Gene-Edited Channel Catfish (Ictalurus punctatus) via Zygote Injection of CRISPR/Cas9 System

Khalil, Karim; Elayat, M. A.; Khalifa, Elham A.; Daghash, S. M.; Elaswad, Ahmed; Miller, Michael E.; Abdelrahman, Hisham A.; Ye, Zhi; Odin, Ramjie; Drescher, David; Vo, Khoi; Gosh, Kamal; Bugg, William S.; Robinson, Dalton; Dunham, Rex A.

doi:10.1038/s41598-017-07223-7

Cited by 118 publications

(52 citation statements)

References 79 publications

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“…The system consists of a guide RNA which determines the targeted sequence in the genome and a DNA endonuclease enzyme, Cas9 1 , 2 . Since 2013, many gene editing experiments have been conducted on fish species including gene knockout 3 – 7 and gene insertion or knock-in 8 – 11 . Recently, extensive genomic data for channel catfish ( Ictalurus punctatus ) became available after the complete sequencing of its genome 12 .…”

Section: Introductionmentioning

confidence: 99%

Effects of CRISPR/Cas9 dosage on TICAM1 and RBL gene mutation rate, embryonic development, hatchability and fry survival in channel catfish

Elaswad

Khalil

et al. 2018

Sci Rep

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The current study was conducted to assess the effects of microinjection of different dosages of guide RNA (gRNA)/Cas9 protein on the mutation rate, embryo survival, embryonic development, hatchability and early fry survival in channel catfish, Ictalurus punctatus. Guide RNAs targeting two of the channel catfish immune-related genes, toll/interleukin 1 receptor domain-containing adapter molecule (TICAM 1) and rhamnose binding lectin (RBL) genes, were designed and prepared. Three dosages of gRNA/Cas9 protein (low, 2.5 ng gRNA/7.5 ng Cas9, medium, 5 ng gRNA/15 ng Cas9 and high, 7.5 ng gRNA/22.5 ng Cas9) were microinjected into the yolk of one-cell embryos. Mutation rate increased with higher dosages (p < 0.05). Higher dosages increased the mutation frequency in individual embryos where biallelic mutations were detected. For both genes, microinjection procedures increased the embryo mortality (p < 0.05). Increasing the dosage of gRNA/Cas9 protein increased the embryo mortality and reduced the hatching percent (p < 0.05). Embryonic development was delayed when gRNAs targeting RBL gene were injected. Means of fry survival time were similar for different dosages (p > 0.05). The current results lay the foundations for designing gene editing experiments in channel catfish and can be used as a guide for other fish species.

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Section: Introductionmentioning

confidence: 99%

Effects of CRISPR/Cas9 dosage on TICAM1 and RBL gene mutation rate, embryonic development, hatchability and fry survival in channel catfish

Elaswad

Khalil

et al. 2018

Sci Rep

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“…As discussed above for mammals, myostatin has also proven a popular target in fish. Knockout of myostatin a (MSTNa) in channel catfish [42] or myostatin ba (MSTNba) in common carp [43] resulted in an enhanced growth rate, with corresponding increases in size and number of muscle fibers. It should be noted that while mammals have a single MSTN gene, tetraploid species such as the carp have a duplication(s) resulting in two or more copies of myostatin.…”

Section: Aquaculturementioning

confidence: 99%

Livestock breeding for the 21st century: the promise of the editing revolution

Proudfoot¹,

McFarlane²,

Whitelaw³

et al. 2020

Front. Agr. Sci. Eng.

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In recent years there has been a veritable explosion in the use of genome editors to create sitespecific changes, both in vitro and in vivo, to the genomes of a multitude of species for both basic research and biotechnology. Livestock, which form a vital component of most societies, are no exception. While selective breeding has been hugely successful at enhancing some production traits, the rate of progress is often slow and is limited to variants that exist within the breeding population. Genome editing provides the potential to move traits between breeds, in a single generation, with no impact on existing productivity or to develop de novo phenotypes that tackle intractable issues such as disease. As such, genome editors provide huge potential for ongoing livestock development programs in light of increased demand and disease challenge. This review will highlight some of the more notable agricultural applications of this technology in livestock.

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“…The gene editing technology can be used to produce agricultural products in accordance with the need of humans. For example, crops were produced with high yield and resistant to diseases, insects, herbicides, and harsh environment [18,19]; domesticated animals (pig, buffalo) were produced with double muscle phenotype [20]; and aquatic products (catfish) were produced with high level myostatin (MSTN) gene expression [21].…”

Section: Agricultural Productsmentioning

confidence: 99%

Introductory Chapter: Gene Editing Technologies and Applications

Chen¹

2019

Gene Editing - Technologies and Applications

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Generation of Myostatin Gene-Edited Channel Catfish (Ictalurus punctatus) via Zygote Injection of CRISPR/Cas9 System

Cited by 118 publications

References 79 publications

Effects of CRISPR/Cas9 dosage on TICAM1 and RBL gene mutation rate, embryonic development, hatchability and fry survival in channel catfish

Effects of CRISPR/Cas9 dosage on TICAM1 and RBL gene mutation rate, embryonic development, hatchability and fry survival in channel catfish

Livestock breeding for the 21st century: the promise of the editing revolution

Introductory Chapter: Gene Editing Technologies and Applications

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