CRISPR/Cas9-mediated mosaic mutation of <i>SRY</i> gene induces hermaphroditism in rabbits

Song, Yuning; Xu, Yuanyuan; Liang, Mingming; Zhang, Yuxin; Chen, Mao; Deng, Jichao; Li, Zhanjun

doi:10.1042/bsr20171490

Cited by 14 publications

(13 citation statements)

References 23 publications

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“…We demonstrate the application of CRISPR/Cas9 in a mosaic analysis of mosGILT functions in A. gambiae. Genome editing in embryos using CRISPR/Cas9 often results in genetic mosaicism in founder animals (F 0 ), which can be used as models for phenotyping especially when a homozygous gene knockout is developmentally lethal (Mianné et al, 2017;Port et al, 2014;Song et al, 2018). A previous study of CRISPR tools in Drosophila has shown that vasa-Cas9 in combination with U6-gRNA allows efficient biallelic gene disruption, which could readily reveal null mutant phenotypes in the soma (Port et al, 2014).…”

Section: Discussionmentioning

confidence: 99%

Disruption of mosGILT in Anopheles gambiae impairs ovarian development and Plasmodium infection

Yang

Schleicher

Dong

et al. 2019

Journal of Experimental Medicine

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Plasmodium infection in Anopheles is influenced by mosquito-derived factors. We previously showed that a protein in saliva from infected Anopheles, mosquito gamma-interferon–inducible lysosomal thiol reductase (mosGILT), inhibits the ability of sporozoites to traverse cells and readily establish infection of the vertebrate host. To determine whether mosGILT influences Plasmodium within the mosquito, we generated Anopheles gambiae mosquitoes carrying mosaic mutations in the mosGILT gene using CRISPR/CRISPR associated protein 9 (Cas9). Here, we show that female mosaic mosGILT mutant mosquitoes display defects in ovarian development and refractoriness to Plasmodium. Following infection by either Plasmodium berghei or Plasmodium falciparum, mutant mosquitoes have significantly reduced oocyst numbers as a result of increased thioester-containing protein 1 (TEP1)–dependent parasite killing. Expression of vitellogenin (Vg), the major yolk protein that can reduce the parasite-killing efficiency of TEP1, is severely impaired in mutant mosquitoes. MosGILT is a mosquito factor that is essential for ovarian development and indirectly protects both human and rodent Plasmodium species from mosquito immunity.

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Section: Discussionmentioning

confidence: 99%

Disruption of mosGILT in Anopheles gambiae impairs ovarian development and Plasmodium infection

Yang

Schleicher

Dong

et al. 2019

Journal of Experimental Medicine

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“…The gene-edited rabbits were successfully created in previous research by our group. 11 , 12 , 13 , 14 The protocol for mutation detection has been described previously. 17 DNA from knockout rabbits was isolated using the TIANamp genomic DNA kit (Tiangen, Beijing, China) according to the manufacturer’s instructions.…”

Section: Methodsmentioning

confidence: 99%

Large-Fragment Deletions Induced by Cas9 Cleavage while Not in the BEs System

Song

Liu

Zhang

et al. 2020

Molecular Therapy - Nucleic Acids

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CRISPR-Cas9 and base editors (BEs) systems are poised to become the gene-editing tool of choice in clinical contexts; however, large-fragment deletion was found in Cas9-mediated mutation cells and mice. In this study, by analyzing 16 gene-edited rabbit lines (including 112 rabbits) generated using SpCas9, BEs, xCas9, and xCas9-BEs with long-range PCR genotyping and long-read sequencing by the PacBio platform, we show the extension of thousands of base fragment deletions in single-guide RNA/Cas9 and xCas9 system mutation rabbits, but no deletions were found in BE-induced mutation rabbits. Thus, we first validated that no large-fragment deletion was induced by the BEs system, suggesting that BE systems can be beneficial tools for the further development of highly accurate and secure gene therapy for the clinical treatment of human genetic disorders.

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“…To determine whether chimeric mutations in the SRY gene induced hermaphroditism, the internal genitalia of SRY-mutant chimeric rabbits were also analyzed. The generation of the model provided a platform to understand the pathogenesis of hermaphroditism and identify novel therapies for human clinical hermaphroditism with male-to-female sex reversal syndrome (43).…”

Section: Sex Selection Strategies During Spermatogenesismentioning

confidence: 99%

Sex Manipulation Technologies Progress in Livestock: A Review

Xie

et al. 2020

Front. Vet. Sci.

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Sex manipulation technologies allow predetermination of the sex of animal offspring by altering the normal reproductive process. In livestock production, the difference in type and gender can translate into significant economic benefits, including alleviation of severe food shortages. In livestock, however, the commercial application of sex manipulation technologies is currently available for cattle only. In this review, we described the brief history of sex manipulation, and the research progresses of common methods used in sex manipulation thus far. Information presented in this review can inform future studies on expanding the scope and use of sex manipulation technologies in livestock.

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CRISPR/Cas9-mediated mosaic mutation of SRY gene induces hermaphroditism in rabbits

Abstract: Hermaphroditism is a rare disorder that affects sexual development, resulting in

Cited by 14 publications

References 23 publications

Disruption of mosGILT in Anopheles gambiae impairs ovarian development and Plasmodium infection

Disruption of mosGILT in Anopheles gambiae impairs ovarian development and Plasmodium infection

Large-Fragment Deletions Induced by Cas9 Cleavage while Not in the BEs System

Sex Manipulation Technologies Progress in Livestock: A Review

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