Abstract CRISPR-based gene drives can spread through wild populations by biasing their own transmission above the 50% value predicted Mendelian inheritance. These technologies offer population-engineering solutions for combating vector-borne diseases, managing crop pests, and supporting ecosystem conservation efforts. Current raise safety concerns unintended propagation. Herein, we address such splitting drive components, Cas9 gRNAs, into separate alleles to form a trans-complementing...

One method for reducing the impact of vector-borne diseases is through use CRISPR-based gene drives, which manipulate insect populations due to their ability rapidly propagate desired genetic traits into a target population. However, all current drives employ Cas9 nuclease that constitutively active, impeding our control over propagation abilities and limiting generation alternative drive arrangements. Yet, other nucleases such as temperature sensitive Cas12a have not been explored designs...

Synthetic CRISPR-based gene-drive systems have tremendous potential in public health and agriculture, such as for fighting vector-borne diseases or suppressing crop pest populations. These elements can rapidly spread a population by breaching the inheritance limit of 50% dictated Mendel's law gene segregation, making them promising tool engineering. However, current technologies lack control over their propagation capacity, there are important concerns about unchecked spreading. Here, we...

Abstract Culex mosquitoes are a global vector for multiple human and animal diseases, including West Nile virus, lymphatic filariasis, avian malaria, posing constant threat to public health, livestock, companion animals, endangered birds. While rising insecticide resistance has threatened the control of mosquitoes, advances in CRISPR genome-editing tools have fostered development alternative genetic strategies such as gene drive systems fight disease vectors. However, though gene-drive...

One of the genes involved in Charcot–Marie–Tooth (CMT) disease, an inherited peripheral neuropathy, is GDAP1. In this work, we show that there a true ortholog gene Drosophila, which have named Gdap1. By up- and down-regulation Gdap1 tissue-specific manner, altering its levels expression produces changes mitochondrial size, morphology distribution, neuronal muscular degeneration. Interestingly, degeneration tissue-autonomous not dependent on innervation. Metabolic analyses our experimental...

Repair of double-strand breaks (DSBs) in somatic cells is primarily accomplished by error-prone nonhomologous end joining and less frequently precise homology-directed repair preferentially using the sister chromatid as a template. Here, Drosophila system performs efficient both DSBs single-strand (SSBs) intact sequences from homologous chromosome process we refer to chromosome-templated (HTR). Unexpectedly, HTR-mediated allelic conversion at white locus was more (40 65%) response SSBs...

Homing CRISPR gene drives could aid in curbing the spread of vector-borne diseases and controlling crop pest invasive species populations due to an inheritance rate that surpasses Mendelian laws. However, this technology suffers from resistance alleles formed when drive-induced DNA break is repaired by error-prone pathways, which creates mutations disrupt gRNA recognition sequence prevent further gene-drive propagation. Here, we attempt counteract encoding additional gRNAs target most...

Members of the Junctophilin (JPH) protein family have emerged as key actors in all excitable cells, with crucial implications for human pathophysiology. In mammals, this consists four members (JPH1-JPH4) that are differentially expressed throughout cells. The analysis knockout mice lacking JPH subtypes has demonstrated their essential contribution to physiological functions skeletal and cardiac muscles neurons. Moreover, mutations JPH2 gene associated hypertrophic dilated cardiomyopathies;...

CRISPR base editors can introduce point mutations into DNA precisely, and cytosine (CBEs) catalyze C to T transitions. While CBEs have been thoroughly explored in cell culture organisms such as mice, little is known about editing insects. In this study, we evaluated germline rates of three different expressed under actin (ubiquitous) or nanos (germline) promoters utilizing Drosophila melanogaster. The original Rattus norvegicus-derived deaminase APOBEC1 (rAPO-1) displayed high (~99%) with...

CRISPR-based gene-drives have been proposed for managing insect populations, including disease-transmitting mosquitoes, due to their ability bias inheritance toward super-Mendelian rates (>50%). Current technologies use a Cas9 that introduces DNA double-strand breaks into the opposing wild-type allele replace it with copy of gene-drive via homology-directed repair. However, different versions is unexplored, and alternative approaches could increase available toolkit designs. Here, we report...

ABSTRACT By surpassing the 50% inheritance limit of Mendel’s law independent assortment, CRISPR-based gene drives have potential to fight vector-borne diseases or suppress crop pests. However, contemporary could spread unchecked, posing safety concerns that their use in both laboratory and field settings. Current technologies also lack chemical control strategies, which be applied for dose, spatial temporal drives. We describe Drosophila first gene-drive system controlled by an engineered...

ABSTRACT CRISPR-based gene drives spread through populations bypassing the dictates of Mendelian genetics, offering a population-engineering tool for tackling vector-borne diseases, managing crop pests, and helping island conservation efforts; unfortunately, current technologies raise safety concerns unintended propagation. Herein, we address this by splitting two drive components, Cas9 gRNAs, into separate alleles to form novel trans-complementing split–gene-drive (tGD) demonstrate its...

In a global aging population, it is important to understand the factors affecting systemic and lifespan. Mitohormesis, an adaptive response caused by different insults mitochondrial network, triggers from nuclear genome inducing several pathways that promote longevity metabolic health. Understanding role of function during process could help biomarker identification development novel strategies for healthy aging. Herein, we interfered muscle expression Drosophila genes Marf Opa1, two encode...

Due to limitations in conventional disease vector control strategies including the rise of insecticide resistance natural populations mosquitoes, genetic using CRISPR gene drive systems have been under serious consideration. The identification target sites mosquito is a key aspect for developing efficient strategies. While genome-wide Cas9 explored precise evaluation focused on coding sequence (CDS) lacking. Additionally, site polymorphisms not characterized other nucleases such as Cas12a,...

ABSTRACT One method for reducing the impact of vector-borne diseases is through use CRISPR-based gene drives, which manipulate insect populations due to their ability rapidly propagate desired genetic traits into a target population. However, all current drives employ Cas9 nuclease that constitutively active, impeding our control over propagation abilities and limiting generation novel drive arrangements. Yet, other nucleases such as temperature-sensitive Cas12a have not been explored...

ABSTRACT Culex mosquitoes are a global vector for multiple human and animal diseases, including West Nile virus, lymphatic filariasis, avian malaria, posing constant threat to public health, livestock, companion animals, endangered birds. While rising insecticide resistance has threatened the control of mosquitoes, advances in CRISPR genome-editing tools have fostered development alternative genetic strategies such as gene drive systems fight disease vectors. However, though gene-drive...

ABSTRACT CRISPR-based gene-drive systems have been proposed for managing insect populations, including disease-transmitting mosquitoes, due to their ability bias inheritance towards super-Mendelian rates (>50%). Current technologies employ a Cas9 that introduces DNA double-strand breaks into the opposing wildtype allele replace it with copy of gene drive via homology-directed repair. Yet, use different Cas9s versions is unexplored, and alternative approaches could increase available...