Masculinization of Zebrafish Through Partial Depletion of Primordial Germ Cells by Injecting Diluted Morpholino Oligonucleotides into Embryos

Orbán, László; Saju, Jolly M.; Tzung, Keh-Weei; Liew, Woei Chang

doi:10.1007/978-1-0716-0970-5_5

Cited by 1 publication

(1 citation statement)

References 24 publications

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“…For general applications, its success depends not only on the transplantation of sufficient numbers of PGCs into the hosts but also on the efficient removal of host PGCs by morpholino-mediated knockdown. Since there are only four to eight PGCs located as four clusters at the blastoderm margin of blastula stage embryos [ 24 ], and reduced numbers of PGCs often lead to the masculinization of zebrafish [ 25 ], it is sometimes challenging to obtain adult females carrying transplanted mutant PGCs.…”

Section: Strategies To Generate Maternal Mutants For Zygotic Lethal Genes In Zebrafishmentioning

confidence: 99%

Circumventing Zygotic Lethality to Generate Maternal Mutants in Zebrafish

Shi

2022

Biology

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Maternal gene products accumulated during oogenesis are essential for supporting early developmental processes in both invertebrates and vertebrates. Therefore, understanding their regulatory functions should provide insights into the maternal control of embryogenesis. The CRISPR/Cas9 genome editing technology has provided a powerful tool for creating genetic mutations to study gene functions and developing disease models to identify new therapeutics. However, many maternal genes are also essential after zygotic genome activation; as a result, loss of their zygotic functions often leads to lethality or sterility, thus preventing the generation of maternal mutants by classical crossing between zygotic homozygous mutant adult animals. Although several approaches, such as the rescue of mutant phenotypes through an injection of the wild-type mRNA, germ-line replacement, and the generation of genetically mosaic females, have been developed to overcome this difficulty, they are often technically challenging and time-consuming or inappropriate for many genes that are essential for late developmental events or for germ-line formation. Recently, a method based on the oocyte transgenic expression of CRISPR/Cas9 and guide RNAs has been designed to eliminate maternal gene products in zebrafish. This approach introduces several tandem guide RNA expression cassettes and a GFP reporter into transgenic embryos expressing Cas9 to create biallelic mutations and inactivate genes of interest specifically in the developing oocytes. It is particularly accessible and allows for the elimination of maternal gene products in one fish generation. By further improving its efficiency, this method can be used for the systematic characterization of maternal-effect genes.

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Section: Strategies To Generate Maternal Mutants For Zygotic Lethal Genes In Zebrafishmentioning

confidence: 99%

Circumventing Zygotic Lethality to Generate Maternal Mutants in Zebrafish

Shi

2022

Biology

View full text Add to dashboard Cite

show abstract

Masculinization of Zebrafish Through Partial Depletion of Primordial Germ Cells by Injecting Diluted Morpholino Oligonucleotides into Embryos

Cited by 1 publication

References 24 publications

Circumventing Zygotic Lethality to Generate Maternal Mutants in Zebrafish

Circumventing Zygotic Lethality to Generate Maternal Mutants in Zebrafish

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