CRISPR/Cas9-mediated knockout of two eye pigmentation genes in the brown planthopper, Nilaparvata lugens (Hemiptera: Delphacidae)

Xue, Wenhua; Xu, Nan; Yuan, Xiaobo; Chen, Haohao; Zhang, Jinli; Fu, Sheng-Jie; Zhang, Chuan-Xi; Xu, Haijun

doi:10.1016/j.ibmb.2017.12.003

Cited by 100 publications

(112 citation statements)

References 39 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…We synthesized two sgRNAs, named BdWhite‐g1 and BdWhite‐g2 , which are located in exons 1 and 3 respectively, and we injected a mixture of Cas9 protein and sgRNA, transcribed in vitro , into fresh eggs that resulted in highly efficient editing of the BdWhite locus. We observed a 100% editing frequency in the surviving flies of the BdWhite‐g2 ‐injected generation; this 100% germline transmission rate was higher than reported for other insects (Wang et al ., ; Xue et al ., ; Chen et al ., ), including B. dorsalis (Zhao et al ., ; Zheng et al ., ; Sim et al ., ). The high mutation efficiency may be due to the high concentration of sgRNA (500 ng/μl) used in this study, and rapid injection of the embryos following oviposition (within 1 h) and the injection of cas9 protein into embryos, rather than cas9 mRNA or plasmid (Zhao et al ., ; Zheng et al ., ).…”

Section: Discussionmentioning

confidence: 99%

“…The CRISPR/Cas9 system is a simple and efficient genome-editing technique that shows significant potential for genetic editing and exploring gene function in nonmodel animals (Choo et al, 2017;Wang et al, 2017;Xue et al, 2017;Chen et al, 2018). The system has been used to edit genomes of several Tephritidae, including B. dorsalis Zhao et al, 2018;Zheng et al, 2018;Li and Handler, 2019;Sim et al, 2019), and here we successfully applied it to study the function of the white gene in B. dorsalis.…”

Section: Discussionmentioning

confidence: 99%

See 1 more Smart Citation

CRISPR/Cas9‐mediated knockout of the eye pigmentation gene white leads to alterations in colour of head spots in the oriental fruit fly, Bactrocera dorsalis

Bai

Zeng

et al. 2019

Insect Molecular Biology

View full text Add to dashboard Cite

The intensely studied white gene is widely used as a genetic marker in Drosophila melanogaster. Here, we cloned and characterized the white gene in an important pest of the fruit industry, Bactrocera dorsalis, to understand its functional role in pigmentation. We obtained BdWhite knockout strains, based on the wild‐type strain, using the CRISPR/Cas9 genome editing system, and found that mutants lost pigmentation in the compound eye and their black head spots. We then examined differences in the expression levels of genes associated with melanin pigmentation between mutants and the wild‐type strain using quantitative reverse transcription PCR. We found that transcription levels of the Bd‐yellow1 were lower in the head of mutants than in the wild‐type strain, and there were no significant differences in expression of the other six genes between mutants and the wild type. Since yellow is critical for melanin biosynthesis (Heinze et al., Scientific Reports. 2017;7:4582), the lower levels of expression of Bd‐yellow1 in mutants led to reduced dark pigmentation in head spots. Our results provide the first evidence, to our knowledge, that white may play a functional role in cuticle pigmentation by affecting the expression of yellow.

show abstract

Section: Discussionmentioning

confidence: 99%

Section: Discussionmentioning

confidence: 99%

CRISPR/Cas9‐mediated knockout of the eye pigmentation gene white leads to alterations in colour of head spots in the oriental fruit fly, Bactrocera dorsalis

Bai

Zeng

et al. 2019

Insect Molecular Biology

View full text Add to dashboard Cite

show abstract

“…To test the efficiency of our CRISPR-Cas9 based genome editing platform in C. quinquefasciatus, we targeted the recessive white (w) gene (CPIJ005542), which codes a protein critical for eye pigment transport. In other species, biallelic mutations in the w gene disrupt production of dark eye pigmentation and generate an easily screenable unpigmented eye colour (Bassett et al, 2014;Ren et al, 2014;Li et al, 2017aLi et al, , 2018Xue et al, 2018). Consequently, we designed three sgRNAs targeting three conserved regions of the third exon of the w gene ( Fig.…”

Section: Mutagenesis Of the White Gene Locus In C Quinquefasciatusmentioning

confidence: 99%

Methods for the generation of heritable germline mutations in the disease vector Culex quinquefasciatus using clustered regularly interspaced short palindrome repeats‐associated protein 9

Liu

et al. 2019

Insect Molecular Biology

View full text Add to dashboard Cite

Culex quinquefasciatus is a vector of many diseases that adversely impact human and animal health; however, compared to other mosquito vectors limited genome engineering technologies have been characterized for this vector. Clustered regularly interspaced short palindrome repeats‐associated protein 9 (CRISPR‐Cas9) based technologies are a powerful tool for genome engineering and functional genetics and consequently have transformed genetic studies in many organisms. Our objective was to improve upon the limited technologies available for genome editing in C. quinquefasciatus to create a reproducible and straightforward method for CRISPR‐Cas9‐targeted mutagenesis in this vector. Here we describe methods to achieve high embryo survival and mutagenesis rates and we provide details on the injection supplies and procedures, embryo handling and guide RNA (gRNA) target designs. Through these efforts, we achieved embryo survival rates and germline mutagenesis rates that greatly exceed previously reported rates in this vector. This work is also the first to characterize the white gene marker in this species, which is a valuable phenotypic marker for future transgenesis or mutagenesis of this vector. Overall, these tools provide the framework for future functional genetic studies in this important disease vector and may support the development of future gene drive and genetic technologies that can be used to control this vector.

show abstract

“…More recently, developmental biologists have used ommochromes of butterfly wings to tackle the mechanisms and the diversity of colour patterning (Sekimura & Nijhout, ). Today, ommochromes are also involved in transcriptomic studies of grasshoppers, butterflies, damselflies and spiders (Croucher et al, ; Chauhan et al, ; Connahs, Rhen & Simmons, ; Qiu et al, ; Wang et al, ), as well as in clustered regular interspaced short palindromic repeats/CRISPR‐associated protein 9 (CRISPR/Cas9)‐mediated genome editing (Khan, Reichelt & Heckel, ; Xue et al, ; Zhang & Reed, ). Unfortunately, biochemical knowledge did not keep pace with this increase in functional studies.…”

Section: Introductionmentioning

confidence: 99%

Ommochromes in invertebrates: biochemistry and cell biology

2018

View full text Add to dashboard Cite

Ommochromes are widely occurring coloured molecules of invertebrates, arising from tryptophan catabolism through the so-called Tryptophan → Ommochrome pathway. They are mainly known to mediate compound eye vision, as well as reversible and irreversible colour patterning. Ommochromes might also be involved in cell homeostasis by detoxifying free tryptophan and buffering oxidative stress. These biological functions are directly linked to their unique chromophore, the phenoxazine/phenothiazine system. The most recent reviews on ommochrome biochemistry were published more than 30 years ago, since when new results on the enzymes of the ommochrome pathway, on ommochrome photochemistry as well as on their antiradical capacities have been obtained. Ommochromasomes are the organelles where ommochromes are synthesised and stored. Hence, they play an important role in mediating ommochrome functions. Ommochromasomes are part of the lysosome-related organelles (LROs) family, which includes other pigmented organelles such as vertebrate melanosomes. Ommochromasomes are unique because they are the only LRO for which a recycling process during reversible colour change has been described. Herein, we provide an update on ommochrome biochemistry, photoreactivity and antiradical capacities to explain their diversity and behaviour both in vivo and in vitro. We also highlight new biochemical techniques, such as quantum chemistry, metabolomics and crystallography, which could lead to major advances in their chemical and functional characterisation. We then focus on ommochromasome structure and formation by drawing parallels with the well-characterised melanosomes of vertebrates. The biochemical, genetic, cellular and microscopic tools that have been applied to melanosomes should provide important information on the ommochromasome life cycle. We propose LRO-based models for ommochromasome biogenesis and recycling that could be tested in the future. Using the context of insect compound eyes, we finally emphasise the importance of an integrated approach in understanding the biological functions of ommochromes.

show abstract

CRISPR/Cas9-mediated knockout of two eye pigmentation genes in the brown planthopper, Nilaparvata lugens (Hemiptera: Delphacidae)

Cited by 100 publications

References 39 publications

CRISPR/Cas9‐mediated knockout of the eye pigmentation gene white leads to alterations in colour of head spots in the oriental fruit fly, Bactrocera dorsalis

CRISPR/Cas9‐mediated knockout of the eye pigmentation gene white leads to alterations in colour of head spots in the oriental fruit fly, Bactrocera dorsalis

Methods for the generation of heritable germline mutations in the disease vector Culex quinquefasciatus using clustered regularly interspaced short palindrome repeats‐associated protein 9

Ommochromes in invertebrates: biochemistry and cell biology

Contact Info

Product

Resources

About