The Trithorax-like (Trl) gene of Drosophila melanogaster encodes the multifunctional protein GAGA involved in many cellular processes. We have isolated and described a new hypomorphic mutation of The Trl gene—Trl _en82 . The mutation is the insertion of a 1.4 kb P-element into the 5′ untranslated region. Trl expression decreased in the ovaries of mutant flies by about 30%; however, it caused abnormalities. The Trl _en82 mutation combined with the null allele of Trl caused female sterility: the females laid a few small eggs with abnormal shape. Many egg chambers demonstrated abnormalities in the Trl _en82 mutants: the oocyte had a regular shape and intruded into the egg chamber region with nurse cells; the rapid transport of nurse cell cytoplasm into the oocyte was disturbed, which resulted in the “dumpless” phenotype of the chambers in mutants; follicular cells often did not completely cover the oocyte and concentrated on its posterior end; and the migration of centripetal cells was affected. We propose that the sterility of the Trl _en82 females is due to the abnormal functioning of follicular cells resulting from low Trlexpression. This proposal is confirmed by normalizing the mutant phenotype of Trl _en82 females after the transfection of Trl cDNA. Note that even an insignificant decrease in Trl expression in such females seriously affected the somatic cell functioning, while a significant decrease in its expression in strong hypomorphic mutants affected both somatic and germline cells in the egg chambers.

The regulatory region of the Trl gene was analyzed using the mutation Trl ³⁶⁰⁹ , resulting from the insertion of the P-element into the promoter region of the gene as well as mutations obtained on its basis. It is shown that two last transcription start sites, which are most often used in vitro in S2 cells, are almost not used in vivo. Experimental data indicate that transcription terminators in transposons play an important role in the decrease in the transcription level of the recipient gene.

At present, there are a variety of different approaches to the targeted regulation of gene expression. However, most approaches are devoted to the activation of gene transcription, and the methods for gene silencing are much fewer in number. In this review, we describe the main systems used for the targeted suppression of gene expression (including RNA interference (RNAi), chimeric transcription factors, chimeric zinc finger proteins, transcription activator-like effectors (TALEs)-based repressors, optogenetic tools, and CRISPR/Cas-based repressors) and their application in eukaryotes—plants and animals. We consider the advantages and disadvantages of each approach, compare their effectiveness, and discuss the peculiarities of their usage in plant and animal organisms. This review will be useful for researchers in the field of gene transcription suppression and will allow them to choose the optimal method for suppressing the expression of the gene of interest depending on the research object.

Drosophila has provided a highly attractive model system for studying various tissue- and stage-specific processes as well as their pathologies, including a range of human diseases. The existence of a large number of diverse Gal4 drivers to precisely control the expression patterns of UAS transgenes simplifies such studies. However, the choice of driver is always critical, as its possible ectopic expression in non-target cells and tissues can directly impact the results. Therefore, it is very important to thoroughly characterize both the molecular nature and expression pattern of each Gal4 driver line. Here, we aim to fill such gaps regarding the AB1-Gal4 driver, which is typically used to express UAS transgenes in larval salivary glands. In this fly line, the P{GawB} enhancer trap construct encoding the Gal4 protein resides within overlapping evolutionary conserved spastin (spas) and Mitochondrial Rho (Miro) genes. Both these genes are expressed in a number of tissues, including the central nervous system (CNS), and their human orthologs are associated with neurodegenerative diseases. Consistently, we demonstrate that, in third-instar larvae, the expression pattern of AB1-Gal4 is also not restricted to salivary glands. We detect its activity in a subset of Elav-positive neurons in the CNS, including motor neurons, as well as in specific photoreceptor cells in eye discs.

In plants, the regulation of transgene transcription is typically achieved using chemical agents. A safe alternative to chemically induced systems may be optogenetic systems. The BphP1-QPAS1 system has distinct advantages over other optogenetic systems, as it is activated by near-infrared (NIR, 780 nm) light, which is beyond the spectrum of plant photoreceptors. This system is based on the use of a split transcription factor (TF), consisting of the DNA-binding and dimerization domains of the yeast TF Gal4, fused to the QPAS1 component, along with the transactivation domain VP16 fused to BphP1. Under NIR light, BphP1 interacts with QPAS1, leading to the formation of the functional TF Gal4-VP16. A primary obstacle to using optogenetic systems in plants is their undesired activation under white light, which is vital for normal plant growth. A potential solution to this issue is temporarily removing one component of the split TF from the nucleus under white light. We modified the BphP1-QPAS1 system to activate reporter gene expression in Nicotiana benthamiana leaves using NIR light. We combined BphP1-QPAS1 with several variants of LOV domain-containing proteins activated by blue light (460–480 nm). The best results were achieved by combining the BphP1-QPAS1 system with the AsLOV2 domain, which carries the degron sequence RRRG at the C-terminal Jα helix and initiates the degradation of the chimeric protein NES-Gal4-QPAS1-AsLOV2-RRRG under white light. This modification induced the BphP1-QPAS1 system in tobacco leaves only under NIR light, but not in the dark or under white light. We believe that, in the future, the BphP1-QPAS1 system could be applied to enhance plant resistance to adverse environmental conditions, pests, and viral diseases.

The RCC1 gene is active in ensuring many cellular functions related to cell division in Drosophila melanogaster and Homo sapiens. A detailed comparison of the structure and functions of the RCC1 gene in Drosophila melanogaster and Homo sapiens was carried out using different analytical techniques (bioinformatics, immunofluorescence and confocal microscopy, FISH, and molecular genetic methods). The Drosophila RCC1 gene belongs to the family of housekeeping genes, since it resides in the interbands and gray bands of polytene chromosomes within aquamarine/lazurite chromatin in D. melanogaster. Furthermore, the databases demonstrate that RCC1 in D. melanogaster is expressed in all the tissues at all the developmental stages. According to The Human Protein Atlas, RCC1 in humans also exhibits low tissue specificity for 29 tissues. Immunostaining of polytene chromosomes with RCC1 antibodies revealed approximately 260 sites of RCC1 protein localization exclusively in black bands (sites of developmental genes) and in heterochromatin. The size of the coding gene portions is almost identical for D. melanogaster and H. sapiens, being ~2 kb. The group of Drosophila proteins related to condensed chromatin, RCC1 being a member of this group, has homologs forming similar interaction networks in humans. The conserved nature of the RCC1 gene has been confirmed by cell cycle studies in both species. It was found that expression of the RCC1 gene is upregulated in glioblastoma; the RCC1 protein predominantly resides on centrioles during metaphase.