Transcription activator-like effector nuclease-mediated deletion safely eliminates the major egg allergen ovomucoid in chickens

Ezaki, Ryo; Sakuma, Tetsushi; Kodama, Daisuke; Sasahara, Ryou; Shiraogawa, Taichi; Ichikawa, Kennosuke; Matsuzaki, Mei; Handa, Akihiro; Yamamoto, Takashi; Horiuchi, Hiroyuki

doi:10.1016/j.fct.2023.113703

Cited by 7 publications

(2 citation statements)

References 28 publications

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“…However, there have been relatively few practical conservation initiatives using PGCs for germplasm conservation of indigenous or rare chicken breeds [21][22][23]. In addition, research on the production of genetically modi ed chickens by PGCs using CRISPR/Cas9 and TALEN remains relatively limited [24][25][26][27][28][29][30].…”

Section: Introductionmentioning

confidence: 99%

PGC-based cryobanking, regeneration through germline chimera mating, and CRISPR/Cas9-mediated TYRP1 modification in indigenous Chinese chickens

Wei,

Kinoshita,

Tanabe

et al. 2024

Preprint

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Primordial germ cells (PGCs) are essential precursors for sperm and eggs. PGCs are highly reliable for conserving chicken germplasm and producing genetically modified chickens. Currently, efforts to conserve native chicken germplasm through PGCs and research on genetically modified chickens using CRISPR/Cas9 are limited. In this study, we successfully established 289 PGC lines from eight Chinese chicken populations with an 81.6% derivation rate. Furthermore, we successfully regenerated Piao chickens derived from these PGCs by repropagating cryopreserved PGCs and transplanting them into recipient chickens, resulting in a 12.7% efficiency rate. The regenerated chickens carried mitochondrial DNA from the female PGC donor and the rumplessness (Rp) mutation from male and female PGC donors in a homozygous condition. As a further application, we utilized CRISPR/Cas9 genome editing to disrupt the TYRP1 gene and created KO PGC lines. Transplanting KO cells into male recipients and mating them with wild-type hens enabled the successful generation of four TYRP1 KO chickens. Molecular analysis confirmed a 73 bp deletion in these four chicks, resulting in a frameshift mutation in the TYRP1 target region. These KO chickens had brown plumage due to reduced eumelanin production, observed in both homozygous and hemizygous mutations, compared to wild-type chickens. In conclusion, we have efficiently cultured and cryopreserved chicken PGCs, used them to regenerate chickens, and generated gene-edited chickens with TYRP1 dysfunction. These achievements will greatly impact the conservation of chicken genetic diversity, create new mutation models, and have potential applications in biomedical and avian research and breeding advancements.

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Section: Introductionmentioning

confidence: 99%

PGC-based cryobanking, regeneration through germline chimera mating, and CRISPR/Cas9-mediated TYRP1 modification in indigenous Chinese chickens

Wei,

Kinoshita,

Tanabe

et al. 2024

Preprint

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“…Additionally, the genetic conservation of locally adapted indigenous chicken breeds, which are found globally in every country (Dessie et al, 2011), is important to maintain their genetic diversity and prevent their loss. Furthermore, recent advancements in genome editing technologies allow us to efficiently produce genetically modified chickens, such as those that lay low allergen eggs (Ezaki et al, 2023;Oishi et al, 2016;Park et al, 2014) or possess specific virus resistance (Idoko-Akoh et al, 2023), increasing the importance of biobanking both academic and commercial chicken lines (Fulton & Delany, 2003).…”

Section: Introductionmentioning

confidence: 99%

Innovations in poultry reproduction using cryopreserved avian germ cells

Ichikawa,

McGrew

2024

Reprod Domestic Animals

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Meat and eggs from chicken are the major source of animal protein for the human population. The cryopreservation of poultry species is needed to guarantee sustainable production. Here, we describe the existing cryopreservation technologies for avian reproductive cells using embryonic germ cells, spermatozoa and ovarian tissues. We outline strategies to reconstitute chicken breeds from their cryopreserved embryonic germ cells using surrogate hosts and discuss the perspectives for genetic conservation and reconstitution of chicken and wild avian species using surrogate host animals.

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