The fruits of various pepper cultivars are characterized by a different colour, which is determined by the pigment ratio; carotenoids dominate in ripe fruits, while chlorophylls, in immature fruits. A key regulator of carotenoid biosynthesis is the phytoene synthase encoded by the PSY gene. The Capsicum annuum genome contains two isoforms of this enzyme, localized in leaf (PSY2) and fruit (PSY1) plastids. In this work, the complete PSY1 and PSY2 genes were identified in nine C. annuum cultivars, which differ in ripe fruit colour. PSY1 and PSY2 sequence variability was 2.43 % (69 SNPs) and 1.21 % (36 SNPs). The most variable were PSY1 proteins of the cultivars ‘Maria’ (red-fruited) and ‘Sladkij shokolad’ (red-brown-fruited). All identified PSY1 and PSY2 homologs contained the phytoene synthase domain HH-IPPS and the transit peptide. In the PSY1 and PSY2 HH-IPPS domains, functionally significant sites were determined. For all accessions studied, the active sites (YAKTF and RAYV), aspartate-rich substrate-Mg2+-binding sites (DELVD and DVGED), and other functional residues were shown to be conserved. Transit peptides were more variable, and their similarity in the PSY1 and PSY2 proteins did not exceed 78.68 %. According to the biochemical data obtained, the largest amounts of chlorophylls and carotenoids across the cultivars studied were detected in immature and ripe fruits of the cv. ‘Sladkij shokolad’ and ‘Shokoladnyj’. Also, ripe fruits of the cv. ‘Nesozrevayuschij’ (green-fruited) were marked by significant chlorophyll content, but a minimum of carotenoids. The PSY1 and PSY2 expression patterns were determined in the fruit pericarp at three ripening stages in ‘Zheltyj buket’, ‘Sladkij shokolad’, ‘Karmin’ and ‘Nesozrevayuschij’, which have different ripe fruit colours: yellow, red-brown, dark red and green, respectively. In the leaves of the cultivars studied, PSY1 expression levels varied significantly. All cultivars were characterized by increased PSY1 transcription as the fruit ripened; the maximum transcription level was found in the ripe fruit of ‘Sladkij shokolad’, and the lowest, in ‘Nesozrevayuschij’. PSY2 transcripts were detected not only in the leaves and immature fruits, but also in ripe fruits. Assessment of a possible correlation of PSY1 and PSY2 transcription with carotenoid and chlorophyll content revealed a direct relationship between PSY1 expression level and carotenoid pigmentation during fruit ripening. It has been suggested that the absence of a typical pericarp pigmentation pattern in ‘Nesozrevayuschij’ may be associated with impaired chromoplast formation.

The development of informative polymorphic DNA markers for poorly studied genera is an important step in population analyses of living organisms, including those that play very important ecological roles in harsh environments, such as desert and semi-desert area. Examples of those poorly studied desert species are Agriophyllum squarrosum L. and Agriophyllum minus Fisch. & Mey. However, a recent RNA-sequencing project in A. squarrosum has proposed a large set of hypothetical SSR (simple sequence repeat) markers. In this work, 11 novel polymorphic SSRs were found due to the screening of 24 randomly selected SSRs for three populations of A. squarrosum and one population of A. minus. The analysis of 11 SSRs revealed 16 polymorphic loci in two Agriophyllum species, 8 polymorphic loci within three populations of A. squarrosum, and 6 polymorphic loci in the population of A. minus. Statistical analyses showed high interspecific, but relatively low intraspecific genetic diversity. The phylogenetic clusterization and population structure analysis have demonstrated a clear segregation of A. minus from A. squarrosum, as well as the separation of population 1 from populations 2 and 3 of A. squarrosum. Thus, we identified the set of novel and informative SSR markers suitable for the study of genetic diversity in Agriophyllum.

The review considers quarantine species and nematode pathotypes potentially dangerous for domestic potato production. Potatoes are affected by more than 30 types of parasitic nematodes, but the review focuses on the most harmful representatives of genera that cause great damage to potato production: Globodera, Ditylenchus, Nacob bus and Meloidogyne. Phytopathological and molecular methods of identification of species and pathotypes and the main achievements in studying the population variability of parasitic potato nematodes were analyzed. It was shown that due to the peculiarities of the life cycle of nematodes and lability of their genomes, the genetic variability of these organisms is very high, which creates a threat of forming new pathogenic genotypes of the parasites. The information about the intra- and interpopulation variability of nematodes is important for studying the ways of introduction and distribution of separate species, as well as for searching for the correlations of molecular markers with the pathotype. Phylogenetic studies based on modern data on genetic variability of populations have allowed to reveal species complexes in Globodera pallida (Stone) Behrens and Nacobbus aberrans (Thorne) Thorne & Allen (sensu lato), including cryptic species. The main components of successful protection preventing a wide distribution of parasitic nematodes are quarantine measures, agricultural techniques, biological methods of protection and cultivation of resistant cultivars. Special attention in the review is paid to the breeding of potato cultivars with durable resistance to various nematode pathotypes, because the cultivation of such varieties is the most ecologically safe and economically advantageous way to prevent epiphytoties. Currently, significant progress has been made in the genetic protection of potato cultivars, especially against cyst-forming nematodes. The review provides data on sources of potato resistance to parasitic nematodes identified in collections of wild and cultivated species. Data on identified R-gens and QTL of resistance that have been introduced into breeding varieties using different methods and approaches are analyzed. The literature data on the study of structural and functional organization of genes for resistance to potato cyst nematodes are given. The results of molecular research on revealing the polymorphisms of loci involved in the control of resistance to cyst and gall nematodes, the development of molecular markers of certain genes and their use in marker-assisted selection for developing of new resistant cultivars, including those with group resistance, are considered.

Pyrenophora tritici-repentis is a causative agent of tan spot in wheat. In recent years, there has been an increasing spread and harmfulness of wheat tan spot. The aim of the research was to study the racial composition of the P. tritici-repentis population in the Republic of Kazakhstan. A collection of 30 common wheat accessions, including promising lines and cultivars from Kazakhstan and CIMMYT–ICARDA, was assessed for resistance to P. triticirepentis in a greenhouse and characterized using the Xfcp623 molecular marker, diagnostic for the Tsn1 gene. Monosporic isolates of P. tritici-repentis isolated from the southeastern region were assigned to certain races based on the manifestation of symptoms of necrosis/chlorosis on standard differentials (Glenlea, 6B662, 6B365). Five races of P. tritici-repentis have been identified, including races 1, 2, 3, 7 and 8. It has been shown that races 1 and 8 of P. tritici-repentis are dominant. As a result of the analysis of the frequency of occurrence of the P. tritici-repentis races, it was found that race 1 (50 %) producing Ptr ToxA and Ptr ToxB and race 8 (35 %) producing Ptr ToxA, Ptr ToxB and Ptr ToxC turned out to be dominant. From a practical point of view, of greatest interest are 16 wheat samples, which demonstrated resistance to race 1 and confirmed insensitivity to Ptr ToxA in a molecular screening. These include eight Kazakhstani (4_PSI, 10204_2_KSI, 10204_3_KSI, 10205_2_KSI, 10205_3_KSI, 605_SP2, 632_SP2, Dana) and seven foreign lines (KR11-20, KR11-03, KR11-9014, 11KR-13, KR12-9025, KR12-07, GN-68/2003). The results of this study are of interest in wheat breeding programs for tan spot resistance.

Under climate change, the need for crops resistant to abiotic and biotic stresses increases. Lathyrus spp. Are characterized by a high nutritional value of their green biomass. The grass pea is one of the most resistant to drought, aterlogging, cold, salinity, diseases and pests among cultivated legumes, and it is grown at minimal cost. The development of new Lathyrus cultivars with an improved composition of nutrients will make it possible to produce high-quality animal feed in areas with extremely unstable weather conditions. With this in view, the patterns of variability in the parameters of the carbohydrate complex (sugars, their lactone and methyl forms), polyols (including phenol-containing alcohols), phytosterols, free fatty acids (FFA) and acylglycerols were studied in the green biomass of 32 accessions representing Lathyrus sativus L., L. tuberosus L., L. sylvestris L., L. vernus (L.) Bernh., L. latifolius L. and L. linifolius (Reichard) Bassler. from the VIR collection, reproduced in Leningrad oblast under contrasting conditions of 2012 and 2013. The content of identified compounds varied depending on the genotype, species, and weather. High temperatures and high precipitation in 2013 contributed to the accumulation of monosaccharides, and the colder and drier conditions of 2012 to an increase in oligosaccharides, most polyols, and FFA. The cultivated species (L. sativus) was distinguished by its high sugar content, and the wild species as follows: L. latifolius by FFA; L. linifolius by ononitol, myo-inositol, and glycerol 3-phosphate; L. vernus by MAG and methylpentofuranoside. The resulting data showed that the Lathyrus accessions studied are promising for breeding stress-resistant cultivars of high nutritional quality.

The creation of varieties adapted to changing environmental conditions, resistant to various pathogens, and satisfying various grain purposes is impossible without using the genetic diversity of wheat. One of the ways to expand the genetic diversity of wheat is to introduce new variants of genes from the genetic pool of congeners and wild relatives into the genotypes of existing varieties. In this study, we used 10 lines from the Arsenal collection created on the genetic basis of the spring variety ‘Rodina’ and the diploid species Aegilops speltoides in the Federal Research Center “Nemchinovka” in 1994. The lines were previously characterized for the presence of translocations and chromosomal rearrangements cytologically and using molecular markers. Technological analyses were performed on grain obtained in Western Siberia and Moscow region. The aim of this study was to establish the possibilities of expanding the phenotypic diversity for technological properties of grain and flour as a result of such hybridization of bread wheat and the diploid cereal Aegilops speltoides. The variety ‘Rodina’ forms a vitreous grain with a high gluten content in Siberia, but has low physical properties of flour and dough. Five derived lines were found to have significantly higher protein and gluten content in grain. The highest values under both growing conditions were found in lines 73/00i, 82/00i, and 84/00i. Two lines (69/00i and 76/00i) showed a high flour strength and dough elasticity, characterizing the lines as strong and valuable in quality. These lines can be used for baking bread. Line 82/00i inherited from Ae. speltoides a soft-grain endosperm, which indicates the introgression of the Ha-Sp gene, homoeoallelic to the Ha gene of bread wheat, into ‘Rodina’. Flour of this line is suitable for the manufacture of confectionery without the use of technological additives. The lines generally retained their characteristics in different growing conditions. They can be attracted as donors of new alleles of genes that determine the technological properties of grain and resistance to biotic stresses.

One of the main tasks of genetics and animal breeding is the assessment of genetic diversity and the study of genetic relationships between different breeds and populations using molecular genetic analysis methods. We analysed the polymorphism of microsatellites and the information on the state of genetic diversity and the population structure of local breeds in Russia: the Kemerovo, the Berkshire, the Liven, the Mangalitsa, and the Civilian; in the Republic of Belarus: the Large White and the Black-and-White; and in Ukraine: the White Steppe, as well as commercial breeds of imported origin of domestic reproduction: the Large White, the Landrace, and the Duroc. The materials used for this study were the tissue and DNA samples extracted from 1,194 pigs and DNA of the UNU “Genetic material bank of domestic and wild animal species and birds” of the L.K. Ernst Federal Research Center for Animal Husbandry. Polymorphisms of 10 microsatellites (S0155, S0355, S0386, SW24, SO005, SW72, SW951, S0101, SW240, and SW857) were determined according to the previously developed technique using DNA analyser ABI3130xl. To estimate the allele pool of each population, the average number of alleles (NA), the effective number of alleles (NE ) based on the locus, the rarified allelic richness (AR), the observed (HO ) and expected (HE ) heterozygosity, and the fixation index (FIS) were calculated. The degree of genetic differentiation of the breeds was assessed based on the pairwise values of FST and D. The analysis of the allelic and genetic diversity parameters of the local breeds showed that the maximum and minimum levels of polymorphism were observed in pigs of the Ukrainian White Steppe breed (NA = 6.500, NE = 3.709, and AR = 6.020) and in pigs of the Duroc breed (NA = 4.875, NE = 2.119, and AR = 3.821), respectively. The highest level of genetic diversity was found in the Large White breed of the Republic of Belarus (HO = 0.707 and NE = 0.702). The minimum level of genetic diversity was found in pigs of the imported breeds – the Landrace (HO = 0.459, HE = 0.400) and the Duroc (HO = 0.480, HE = 0.469) – indicating a high selection pressure in these breeds. Based on the results of phylogenetic analysis, the genetic origin of Large White pigs, the breeds, from which the Berkshire pigs originated, and the genetic detachment of the Landrace from the Mangalitsa breeds were revealed. The cluster analysis showed a genetic consolidation of the Black-and-White, the Berkshire, and the Mangalitsa pigs. Additionally, the imported breeds with clustering depending on the origin were characterised by a genetic structure different from that of the other breeds. The information obtained from these studies can serve as a guide for the management and breeding strategies of the pig breeds studied, to allow their better use and conservation.

The quantitative trait loci associated with the immune properties of chickens are of interest from the point of view of obtaining animals resistant to infectious agents using marker-assisted selection. In the process of selecting markers for genomic selection in broiler-type chickens, a non-standard genotype frequency of the RACK1 gene allele (SNP Gga_rs15788101) in the B5 line of broiler-type chicken cross Smena 8 was identified and it was suggested that this gene was involved in selection. Therefore, it was decided to investigate the available polymorphisms in the three genes responsible for the IgY titer (DMA, RACK1 and CD1B). Molecular typing of single nucleotide polymorphisms of three loci revealed an approach to fixation of the unfavorable allele of the DMA gene (SNP Gga_rs15788237), an approach to fixation of the unfavorable allele of the RACK1 gene and the prevalence of the favorable CD1B gene allele (SNP Gga_rs16057130). Analysis of the haplotypes revealed a strong linkage disequilibrium of these genes. This suggests that these genes experience selection pressure. Analysis of the protein-coding sequences of the CD1B and DMA genes of various breeds of chickens revealed a negative selection of these genes. In order to understand whether the fixation of the studied alleles is the result of artificial selection of the B5 line of the cross Smena 8, an analysis of similar loci in layer chickens Hisex White was carried out. The frequencies of the alleles at the loci of the CD1B gene (Gga_rs16057130) and the RACK1 gene (Gga_rs15788101) in the Hisex White chicken genome differ from the frequencies of the alleles obtained for chickens of the B5 line of the cross Smena 8. It can be assumed that the fixation of the allele in the DMA gene (SNP Gga_rs15723) is associated with artificial or natural selection, consistent in broilers and layers. Changes in the loci Gga_rs16057130 and Gga_rs15788101 in the B5 line of the Smena 8 chickens are most likely associated with artificial selection of broiler productivity traits, which can subsequently lead to fixation of alleles at these loci. Artificial breeding of chickens leads to degradation of the variability of genes encoding elements of the immune system, which can cause a decrease in resistance to various diseases. The study of the negative impact of selection of economic traits on immunity should provide means to mitigate negative consequences and help find ways to obtain disease-resistant animals.

Assisted reproductive technologies (ART) increasingly occupy the study of human reproduction. In addition, in developed countries they contribute to breeding of more than 50 % of cattle. In the management of collections of genetic lines of laboratory animals, these technologies are obligatory components of cryopreservation and rederivation. ART procedures include the development of early embryos outside the mother’s body and the high probability of incomplete synchronization of the physiological state of the surrogate mother and transplanted embryos. Since all this occurs at the stage of the highest susceptibility of embryos to epigenetic reprogramming, the full cycle of ART and its individual components can lead to stable phenotypic changes in the offspring. Their reality is confirmed by studies of the morphological and functional characteristics of sexually mature offspring of CD1 outbred mice, obtained using different variants of early embryo transplantation. Comparative studies of body mass and body composition, basal glucose level and response to glucose load (glucose-tolerance test – GTT) have been done on sexually mature males and females. Animals were separated in 4 groups according to the variant of embryo transplantation: group (control) – natural mating; group (2cl-bl) – incubation of 2-cell up to blastocysts; group (2cl-2cl) – removal and transplantation of the 2-cell embryo without incubation; group (Bl-bl) removal and transplantation of the blastocysts without incubation. All embryos were transplanted to recipient females of the same line. It was found that sexually mature offspring obtained with all variants of transplantations had a higher relative fat content and, correspondingly, lower lean mass compared to the control. This effect was more pronounced in females than in males. Unlike body compositions, embryo transplantations had a greater effect on basal glucose concentration and GTT in males than in females. In this case, the offspring of the 2cl-2cl and 2cl-bl groups were characterized by a higher tolerance to glucose load (GTT) compared with the control and the Bl-bl group. Stable deviations of body compositions and glucose homeostasis indices detected in experimental groups of progenies indicate the phenotypic significance of the embryo transplantations per se.

The DISC1 (disrupted in sсhizophrenia 1) gene is associated with brain dysfunctions, which are involved in a variety of mental disorders, such as schizophrenia, depression and bipolar disorder. This is the first study to examine the immune parameters in Disc1-Q31L mice with a point mutation in the second exon of the DISC1 gene compared to mice of the C57BL/6NCrl strain (WT, wild type). A flow cytometry assay has shown that intact Disc1-Q31L mice differ from the WT strain by an increase in the percentage of CD3+ T cells, CD3+CD4+ Т helper cells and CD3+CD4+CD25+ T regulatory cells and a decrease in CD3+CD8+ T cytotoxic/suppressor cells in the peripheral blood. A multiplex analysis revealed differences in the content of cytokines in the brain structures of Disc1-Q31L mice compared to WT mice. The content of pro-inflammatory cytokines was increased in the frontal cortex (IL-6, IL- 17 and IFNγ) and striatum (IFNγ), and decreased in the hippocampus and hypothalamus. At the same time, the levels of IL-1β were decreased in all structures being examined. In addition, the content of anti-inflammatory cytokines IL-4 was increased in the frontal cortex, while IL-10 amount was decreased in the hippocampus. Immune response to sheep red blood cells analyzed by the number of antibody-forming cells in the spleen was higher in Disc1-Q31L mice at the peak of the reaction than in WT mice. Thus, Disc1-Q31L mice are characterized by changes in the pattern of cytokines in the brain structures, an amplification of the peripheral T-cell link with an increase in the content of the subpopulations of CD3+CD4+ T helpers and CD3+CD4+CD25+ T regulatory cells, as well as elevated immune reactivity to antigen in the spleen.

Lung cancer is one of the most common types of cancer in the world. Although the mechanism of lung cancer is still unknown, a large number of studies have found a link between gene polymorphisms and the risk of lung cancer. The tumor suppressor p53 plays a crucial role in maintaining genomic stability and tumor prevention. MDM2 is a critical regulator of the p53 protein. Despite the importance of p53 pathway in cancer, data on the contribution of SNPs of TP53 (rs1042522) and MDM2 (rs2279744) to the development of lung cancer are very contradictory. A metaanalysis that collects quantitative data from individual studies and combines their results has the advantage of improving accuracy, providing reliable estimates, and resolving those issues in which studies on individual associations are not effective enough. The aim of this study was to determine whether the TP53 (rs1042522) and MDM2 (rs2279744) polymorphisms confer susceptibility to lung cancer. A meta-analysis was conducted on the associations between the TP53 (rs1042522) and MDM2 (rs2279744) polymorphisms and lung cancer. A total of 51 comparison studies including 25,366 patients and 25,239 controls were considered in this meta-analysis. The meta-analysis showed no association between lung cancer and MDM2 (rs2279744) under any model. A noteworthy association of TP53 (rs1042522) with susceptibility to lung cancer in overall pooled subjects was observed under three different models (allele contrast, homozygote contrast (additive) and dominant). Stratification by ethnicity indicated an association between the TP53 (rs1042522) and lung cancer in Asians and Caucasians. This meta-analysis demonstrates that the TP53 (rs1042522), but not MDM2 (rs2279744) polymorphism may confer susceptibility to lung cancer.

Reproductive potential is the most important conditional indicator reflecting the ability of individuals in a population to reproduce, survive and develop under optimal environmental conditions. As for humans, the concept of reproductive potential can include the level of the individual’s mental and physical state, which allows them to reproduce healthy offspring when they reach social and physical maturity. Female reproductive potential has been investigated in great detail, whereas the male reproductive potential (MRP) has not received the equal amount of attention as yet. Therefore, here we focused on the human Y chromosome and found candidate single-nucleotide polymorphism (SNP) markers of MRP. With our development named Web-service SNP_TATA_Z-tester, we examined in silico all 35 unannotated SNPs within 70-bp proximal promoters of the three Y-linked genes, CDY2A, SHOX and ZFY, which represent all types of human Y-chromosome genes, namely: unique, pseudo-autosomal, and human X-chromosome gene paralogs, respectively. As a result, we found 11 candidate SNP markers for MRP, which can significantly alter the TATA-binding protein (TBP) binding affinity for promoters of these genes. First of all, we selectively verified in vitro the values of the TBP-promoter affinity under this study, Pearson’s linear correlation between predicted and measured values of which were r = 0.94 (significance p < 0.005). Next, as a discussion, using keyword search tools of the PubMed database, we found clinically proven physiological markers of human pathologies, which correspond to a change in the expression of the genes carrying the candidate SNP markers predicted here. These were markers for spermatogenesis disorders (ZFY: rs1388535808 and rs996955491), for male maturation arrest (CDY2A: rs200670724) as well as for disproportionate short stature at Madelung deformity (e. g., SHOX: rs1452787381) and even for embryogenesis disorders (e. g., SHOX: rs28378830). This indicates a wide range of MRI indicators, alterations in which should be expected in the case of SNPs in the promoters of the human Y-chromosome genes and which can go far beyond changes in male fertility.

The objects of study are the territory of the districts of the West Kazakhstan region of the Republic of Kazakhstan adjacent to the Kapustin Yar test site: Bokeyordinsky, Zhangalinsky, Zhanibeksky, Kaztalovsky, Akzhayiksky and Syrymsky. The purpose of the work is to conduct a radioecological survey of contaminated areas and to study the content of pollutants in environmental objects, biota (rodents, fish and biosubstrates of domestic animals) by physicochemical methods to assess the risk of the landfill’s impact on biota and humans. This paper presents the results of expeditionary and laboratory research on the pollution of environmental objects in the areas adjacent to the landfill. Indicators of the gamma-survey of the levels of radiation background of the surveyed territories as wellas data on pollution of soil, surface and ground waters, dominant forms of plants, biosubstrates (hair of domestic animals: camel, horse, cow) are given. The generally accepted research methods were used: standard sampling me thods, radiological, atomic adsorption spectrophotometry, cytogenetic (micronucleus) method. A reconnaissance and radioecological examination of environmental objects was carried out using analytical methods, which made it possible to determine the quantitative content of toxic components, the content of priority pollutants and radioactive isotopes. It has been established that the values of the volumetric activity of natural and man-made radionuclides in soil samples, drinking water and biosubstrates (pet hair, human peripheral blood samples) from settlements correspond to the value of the control level for this region. The measurements of radiation activity by gamma radiation showed that along the perimeter of the surveyed territory of the test site and in nearby settlements the radiation level is in the range of 0.06–0.014 μSv/h. A slight excess of the level of radioactivity persists in the area near the fall of missiles in the Bokeyordin region. The investigated regions are characterized by an insignificant level of background radiation, the average DER value for the regions as a whole is 0.14 μSv/h. The absolute maximum, 0.73 μSv/h, was recorded at the points of missile impact in the Kaztal region.

After the genome sequence of SARS-CoV-2 (Severe acute respiratory syndrome-related coronavirus 2) was published and the number of infected people began to increase rapidly, many global companies began to develop a vaccine. Almost all known approaches to vaccine design were applied for this purpose, including inactivated viruses, mRNA and DNA-vaccines, vaccines based on various viral vectors, synthetically generated peptides and recombinant proteins produced in cells of insects and mammals. This review considers one of the promising vaccine platforms based on messenger RNA. Until recent years, mRNA-vaccination was out of practical implementation due to high sensitivity to nuclease degradation and consequent instability of drugs based on mRNA. Latest technological advances significantly mitigated the problems of low immunogenicity, instability, and difficulties in RNA-vaccine delivery. It is worth noting that mRNA-vaccines can efficiently activate both components of the immune system, i. e. T-cell and humoral responses. The essential advantage of mRNA-vaccines includes fast, inexpensive, scalable and uniform production providing a large output of desirable products in vitro. Synthesis and purification processes significantly simplify the process technology of mRNA drugs with injectable purity. Thus, mRNA production via in vitro transcription is more advantageous as compared with DNA-vaccines since it is a chemical process without the use of cells. mRNA techniques make it possible to pass all the phases of vaccine development much faster in comparison with the production of vaccines based on inactivated viruses or recombinant proteins. This property is critically important when designing vaccines against viral pathogens as the main problem of disease control includes a time gap between an epidemic and vaccine development. This paper discusses studies on the development of vaccines against coronaviruses including SARS-CoV-2 with special attention to the mRNA technique.