Ranian Kanwal, Kashif Zahoor Muhammad, Zulhussnain Muhammad, Ahmad Aftab
Department of Zoology, Government College University Faisalabad, Pakistan.
Centre of Department of Biochemistry/US-Pakistan Center for Advance Studies in Agriculture and Food Security (USPCAS-AFS), University of Agriculture Faisalabad, Pakistan.
Saudi J Biol Sci. 2022 Apr;29(4):3015-3022. doi: 10.1016/j.sjbs.2022.01.034. Epub 2022 Jan 25.
is a principal vector for several viruses including dengue virus, chikungunya virus and zika virus. Economic burden of mosquito-borne diseases, relative failure of traditional control strategies and the resistance development against insecticides enforces towards genetic manipulation of . Hence, a key gene () which regulate sex differentiation and alternatively splices to form male and female specific transcripts ( and ). CRISPR/Cas9 technique was employed to sex specifically disrupt the female-specific isoforms, and , both of which were shown to be expressed only in female mosquitoes Targeting of at the developmental stage has resulted in various phenotypic anomalies of adult females. The rate of adult mutation phenotype was recorded between 29 and 37% along with anomalies of wing size, proboscis length and reduction in the sizes of pre-blood-meal and after blood-meal ovaries in and microinjected groups, respectively. These findings can be correlated with reduced fecundity rate of G female, where and groups showed reduction rate in range of 23-31%. Furthermore, hatching inhibition rate of 28 to 36% was also observed in G1 generation when compared to the wildtype. Overall, these results demonstrated that disruption has resulted in multiple female traits disruption including decreased fertility of the female that could directly or indirectly associated with reproduction and its disease transmitting abilities. All these findings suggesting that CRISPR works to alter the developmental pathways as predicted, and therefore this method potentially gives us the basis for the sex-ratio distortion system as genetic control approach for the management of this vector.
是包括登革热病毒、基孔肯雅病毒和寨卡病毒在内的多种病毒的主要传播媒介。蚊媒疾病的经济负担、传统控制策略的相对失败以及对杀虫剂产生的抗性促使人们对[蚊子名称]进行基因操纵。因此,一个关键基因[基因名称]调控性别分化,并通过可变剪接形成雄性和雌性特异性转录本([转录本名称1]和[转录本名称2])。采用CRISPR/Cas9技术特异性地破坏雌性特异性异构体[转录本名称1]和[转录本名称2],这两种异构体仅在雌性蚊子中表达。在发育阶段靶向[基因名称]导致成年雌性出现各种表型异常。在[基因名称1]和[基因名称2]显微注射组中,成年突变表型的发生率分别为29%至37%,同时伴有翅大小、喙长度异常以及血餐前和血餐后卵巢大小减小。这些发现与G代雌性的繁殖力降低相关,其中[基因名称1]和[基因名称2]组的降低率在23%至31%之间。此外,与野生型相比,在G1代中还观察到28%至36%的孵化抑制率。总体而言,这些结果表明[基因名称]的破坏导致了多个雌性性状的破坏,包括雌性生育力下降,这可能直接或间接与繁殖及其疾病传播能力相关。所有这些发现表明,CRISPR如预期那样改变了发育途径,因此这种方法可能为作为该传播媒介基因控制方法的性别比例扭曲系统提供基础。