CRISPR Cas9 mediated knockout of sex determination pathway genes in <i>Aedes aegypti</i>

Zulhussnain, Muhammad; Zahoor, Muhammad Kashif; Ranian, Kanwal; Ahmad, Aftab; Jabeen, Farhat

doi:10.1017/s0007485322000505

Cited by 3 publications

(3 citation statements)

References 31 publications

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“…TALENs has been applied in the silkworm BmBLOS2 gene, resulting in an oily silkworm phenotype and succeeding in B. mori clock gene period e knockout; gene knockout of Ostrinia furnacalis sex pheromone related receptor gene resulted in abnormal mating behavior [30,31]. The CRISPR/Cas system has also been used in D. melanogaster [9], A. aegypti [10], Locusta migratoria [32] and Blattella germanica [33]. However, these techniques have not been applied in M. alternatus.…”

Section: Discussionmentioning

confidence: 99%

“…After innovation of zinc finger nucleases (ZFNs), upon emergence of transcription activator-like effecter nuclease (TALENs) and CRISPR/Cas, targeted genome-editing technology is becoming more efficient, less costly and easier to operate. In addition to common model insects such as D. melanogaster, Culex and Tribolium castaneum, the most updated targeted genome-editing technology, the CRISPR/Cas system, is also widely used in Diptera [9,10], Hymenoptera [11], Hemiptera [12], Lepidoptera [13][14][15] and Orthoptera [16]. Therefore, it is urgent to develop a feasible gene-editing system for M. alternatus and expand the system's prevention methods.…”

Section: Introductionmentioning

confidence: 99%

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RNA Interference-Mediated Knockdown of Tryptophan 2,3-Dioxygenase and Kynurenine-3-Monooxygenase in Monochamus Alternatus: Implications for Insect Control

Zhang

Weng

et al. 2023

Forests

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Monochamus alternatus Hope (Coleoptera: Cerambycidae), an invasive beetle that has caused billions of dollars in economic losses, is a serious pest of Pinus massoniana in many Asian countries. Clarifying the eye pigment gene and its knockdown phenotype of M. alternatus can provide functional gene identification and a marker for screening of gene editing, as well as help develop new control ideas. In this study, we first screened the transcriptome and found one homologous gene of tryptophan 2,3-dioxygenase (TDO) and one of kynurenine-3-monooxygenase (KMO). By measuring the expression levels of TDO and KMO in different developmental periods, it was indicated that TDO and KMO were expressed in various stages of M. alternatus. The gene expression of MaKMO was higher than MaTDO, showing high expression after pupation and decreasing at the beginning of eclosion. MaTDO and MaKMO were knocked down using RNA interference technology in different periods of expression, and the temporal expression changes were obtained using RT-qPCR technology. The results showed that the expressions of MaTDO and MaKMO were significantly inhibited by the injection of dsRNA; the expressions of MaTDO at 48 h, 96 h and after pupation were 21.9%, 32.3% and 59.2%, respectively, meanwhile, those of KMO were 23.4%, 25.0% and 69.7%, respectively. There was a significant change in eye color, and the beetles were able to pupate normally without their activity being affected. Therefore, both MaTDO and MaKMO can be used as tag genes for M. alternatus. A dominant marker system based on eye color can be developed for the genetic manipulation and control of M. alternatus.

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Section: Discussionmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

RNA Interference-Mediated Knockdown of Tryptophan 2,3-Dioxygenase and Kynurenine-3-Monooxygenase in Monochamus Alternatus: Implications for Insect Control

Zhang

Weng

et al. 2023

Forests

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“…CRISPR-Cas9 genome-editing technology is effective at decreasing the sex ratio in mosquito populations. Another study by Zulhusnain et al [81] targeted and attenuated two genes found in Aedes aegypti mosquitoes, namely, doublesex (Aaedsx) and sexlethal (AaeSxl). Inoculation of Aedes aeypti eggs with the Cas9 protein (333 ng µl -1 ) and gRNAs (100 ng µl -1 ) resulted in the generation of the Aaedsx and AaeSxl knockout (KO) strains.…”

Section: Crispr-cas9-based Interventions Have the Potential To Modify...mentioning

confidence: 99%

CRISPR-Cas9 Genome Editing Technology for Zoonotic Disease Control in Indonesia: A Comprehensive Review

Adnyana,

Eljatin,

Sudaryati

et al. 2024

JMHT

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The increasing emergence of zoonotic diseases originating from vectors, rodents, mammals, and others has increased the potential for outbreaks of pandemics in the Indonesian territory. Although control and prevention have been implemented, these efforts have not yet revealed a bright spot; therefore, elaboration with advanced CRISPR-Cas9 technology is a way to accelerate efforts to control zoonotic diseases in Indonesia. However, there is limited literature on this topic. This review aims to comprehensively describe, identify, and summarize the application of CRISPR-Cas9 genome-editing technology in zoonotic disease control in Indonesia. Our findings show that CRISPR-Cas9 genome editing technology offers an innovative approach for zoonotic disease control by targeting disease vectors, modifying animal reservoirs, improving disease surveillance, enhancing vaccine development, and exploring traditional medicine candidates and immunotherapy. The high level of precision, efficiency, and versatility in targeting genomes capable of disrupting, damaging, and disrupting the disease transmission cycle in pathogens makes CRISPR-Cas9 highly effective. However, challenges such as off-target impacts, regulatory complexity, and ethical considerations must be overcome with inter- and multidisciplinary collaboration to promote transparency, equity, and public engagement throughout the process of implementing this technology in the field, especially in Indonesia.

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Using CRISPR-Cas9 to create knockout mutants in insects

Hillary,

Ceasar

2024

CABI Reviews

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Clustered regularly interspaced short palindromic repeats (CRISPR) and CRISPR-associated protein 9 (CRISPR/Cas9) is a potent genome editing tool that has rapidly revolutionized research in natural science disciplines. The CRISPR/Cas9 system surpasses traditional zinc finger nucleases and transcription activator-like effector nucleases because of its simplicity in design, cost-effectiveness, and labor intensity. The CRISPR/Cas9-mediated genome editing system has been used to generate precise and targeted mutations in many different organisms, including insects. However, there is still a lack of a comprehensive review that systematically summarizes the successful utilization of the CRISPR/Cas9 system in generating knockout (KO) mutations in insects (both model and non-model). Thus, this review provides a comprehensive analysis of the current advancements in CRISPR/Cas9-based KO mutants in insects, including both model and non-model species. We also discussed the limitations of the CRISPR/Cas9 system. Finally, we discuss the biosafety concerns associated with the release of CRISPR/Cas9-edited insects into the environment and draw inroads and insights into the future application of the CRISPR/Cas9 system in insects.

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CRISPR Cas9 mediated knockout of sex determination pathway genes in Aedes aegypti

Cited by 3 publications

References 31 publications

RNA Interference-Mediated Knockdown of Tryptophan 2,3-Dioxygenase and Kynurenine-3-Monooxygenase in Monochamus Alternatus: Implications for Insect Control

RNA Interference-Mediated Knockdown of Tryptophan 2,3-Dioxygenase and Kynurenine-3-Monooxygenase in Monochamus Alternatus: Implications for Insect Control

CRISPR-Cas9 Genome Editing Technology for Zoonotic Disease Control in Indonesia: A Comprehensive Review

Using CRISPR-Cas9 to create knockout mutants in insects

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