Complementary Genes Research Articles

In Situ DNA Detection Using a Phase-Demodulated High-Order Intermodal Interferometer Based on an Exposed-Core Microstructure Fiber

In this paper, we demonstrate a phase-demodulated high-order intermodal interference based on exposed-core microstructure fiber (ECF), and it is applied for low concentration in-situ deafness gene DNA detection. The simulation results show that the phase sensitivity of the high-order interference mode (LP <sub xmlns:mml="http://www.w3.org/1998/Math/MathML" xmlns:xlink="http://www.w3.org/1999/xlink">01</sub> -LP <sub xmlns:mml="http://www.w3.org/1998/Math/MathML" xmlns:xlink="http://www.w3.org/1999/xlink">21a</sub> ) is about 6 times higher than that of the low-order (LP <sub xmlns:mml="http://www.w3.org/1998/Math/MathML" xmlns:xlink="http://www.w3.org/1999/xlink">01</sub> -LP <sub xmlns:mml="http://www.w3.org/1998/Math/MathML" xmlns:xlink="http://www.w3.org/1999/xlink">11b</sub> ), which is different from the traditional wavelength coded demodulation method. Hybridization experiments of complementary and non-complementary deafness gene DNA fragments were carried out, and the sensing system was obtained to screen for complementary gene fragments. DNA complementary fragments with concentrations as low as 1 nM can be detected with a corresponding phase variation of 7.32 °. The limit of detection is as low as 0.089 nM. The experiment and theoretical analysis prove that this fiber optic biosensor can obtain high sensitivity and signal-to-noise ratio with phase-demodulation high-order intermodal interference scheme, which provides an alternative for the subsequent studies of high sensitivity and high precision sensors.

Euglycemia is affected by stress defense factor hepatocyte NRF1, but not NRF2

Hepatocyte stress signaling has been established to alter glucose metabolism and impair systemic glucose homeostasis. In contrast, the role of stress defenses in the control of glucose homeostasis is less understood. Nuclear factor erythroid 2 related factor-1 (NRF1) and −2 (NRF2) are transcription factors that promote stress defense and can exert hepatocyte stress defense programming via complementary gene regulation. To identify whether there are independent or complementary roles of these factors in hepatocytes on glucose homeostasis, we investigated the effect of adult-onset, hepatocyte-specific deletion of NRF1, NRF2, or both on glycemia in mice fed 1–3 weeks with a mildly stressful diet enriched with fat, fructose, and cholesterol. Compared to respective control, NRF1 deficiency and combined deficiency reduced glycemia, in some cases resulting in hypoglycemia, whereas there was no effect of NRF2 deficiency. However, reduced glycemia in NRF1 deficiency did not occur in the leptin-deficient mouse model of obesity and diabetes, suggesting hepatocyte NRF1 support defenses that counteract hypoglycemia but does not promote hyperglycemia. Consistent with this, NRF1 deficiency was associated with reduced liver glycogen and glycogen synthase expression as well as marked alteration to circulating level of glycemia-influencing hormones, growth hormone and insulin-like growth factor-1 (IGF1). Overall, we identify a role for hepatocyte NRF1 in modulating glucose homeostasis, which may be linked to liver glycogen storage and the growth hormone/IGF1 axis.

Evolution of Bipartite and Segmented Viruses from Monopartite Viruses.

RNA viruses may be monopartite (all genes on one strand), multipartite (two or more strands packaged separately) or segmented (two or more strands packaged together). In this article, we consider competition between a complete monopartite virus, A, and two defective viruses, D and E, that have complementary genes. We use stochastic models that follow gene translation, RNA replication, virus assembly, and transmission between cells. D and E multiply faster than A when stored in the same host as A or when together in the same host, but they cannot multiply alone. D and E strands are packaged as separate particles unless a mechanism evolves that allows assembly of D + E segmented particles. We show that if defective viruses assemble rapidly into separate particles, the formation of segmented particles is selected against. In this case, D and E spread as parasites of A, and the bipartite D + E combination eliminates A if the transmissibility is high. Alternatively, if defective strands do not assemble rapidly into separate particles, then a mechanism for assembly of segmented particles is selected for. In this case, the segmented virus can eliminate A if transmissibility is high. Conditions of excess protein resources favor bipartite viruses, while conditions of excess RNA resources favor segmented viruses. We study the error threshold behavior that arises when deleterious mutations are introduced. Relative to bipartite and segmented viruses, deleterious mutations favor monopartite viruses. A monopartite virus can give rise to either a bipartite or a segmented virus, but it is unlikely that both will originate from the same virus.

Mapping of leaf rust resistance genes in common wheat ‘Guinong08-6’

Wheat leaf rust (Puccinia triticina Eriks.), a devastating disease of wheat in the world, causes severe yield losses and therefore the development of resistant cultivars is very important. Here, a Chinese wheat line (Guinong08-6) showed adult-plant resistance against mixed fungal isolates of leaf rust, which is common in Guiyang region. It was crossed with a susceptible wheat line (Guinong19) to develop F1, F2, and F3 hybrids. Combined SSR and STS markers were used to map leaf rust resistance genes in Guinong08-6, and the resistance phenotype of Guinong08-6 was co-regulated by two complementary dominant genes, named LrGn08-6A and LrGn08-6B. LrGn08-6A was mapped to chromosome 2AS with markers URIC-LN2 and Xgpw2204, which flanked the gene at distances of 1.8 centimorgan (cM) and 14.83 cM, respectively. LrGn08-6B was mapped to chromosome 4DL with markers Xgpw342 and Xbarc93, which both flanked the gene at a distance of 26.57 cM. Genetic and molecular marker analyses demonstrated that LrGn08-6A, which was inherited from Aegilops ventricosa may be the resistance gene Lr37, while LrGn08-6B may be a newly discovered leaf rust resistance gene.

Complementary gene regulation by NRF1 and NRF2 protects against hepatic cholesterol overload

Hepatic cholesterol overload promotes steatohepatitis. Insufficient understanding of liver stress defense impedes therapy development. Here, we elucidate the role of stress defense transcription factors, nuclear factor erythroid 2 related factor-1 (NRF1) and -2 (NRF2), in counteracting cholesterol-linked liver stress. Using a diet that increases liver cholesterol storage, expression profiles and phenotypes of liver from mice with hepatocyte deficiency of NRF1, NRF2, or both are compared with controls, and chromatin immunoprecipitation sequencing is undertaken to identify target genes. Results show NRF1 and NRF2 co-regulate genes that eliminate cholesterol and mitigate inflammation and oxidative damage. Combined deficiency, but not deficiency of either alone, results in severe steatohepatitis, hepatic cholesterol overload and crystallization, altered bile acid metabolism, and decreased biliary cholesterol. Moreover, therapeutic effects of NRF2-activating drug bardoxolone require NRF1 and are supplemented by NRF1 overexpression. Thus, we discover complementary gene programming by NRF1 and NRF2 that counteract cholesterol-associated fatty liver disease progression.

Sparse representation learning derives biological features with explicit gene weights from the Allen Mouse Brain Atlas.

Unsupervised learning methods are commonly used to detect features within transcriptomic data and ultimately derive meaningful representations of biology. Contributions of individual genes to any feature however becomes convolved with each learning step, requiring follow up analysis and validation to understand what biology might be represented by a cluster on a low dimensional plot. We sought learning methods that could preserve the gene information of detected features, using the spatial transcriptomic data and anatomical labels of the Allen Mouse Brain Atlas as a test dataset with verifiable ground truth. We established metrics for accurate representation of molecular anatomy to find sparse learning approaches were uniquely capable of generating anatomical representations and gene weights in a single learning step. Fit to labeled anatomy was highly correlated with intrinsic properties of the data, offering a means to optimize parameters without established ground truth. Once representations were derived, complementary gene lists could be further compressed to generate a low complexity dataset, or to probe for individual features with >95% accuracy. We demonstrate the utility of sparse learning as a means to derive biologically meaningful representations from transcriptomic data and reduce the complexity of large datasets while preserving intelligible gene information throughout the analysis.

An enhanced network of energy metabolism, lysine acetylation, and growth-promoting protein accumulation is associated with heterosis in elite hybrid rice

Heterosis refers to the superior performance of a hybrid compared with its parental lines. Although several genetic and molecular models have been proposed to explain heterosis, it remains unclear how hybrid cells integrate complementary gene expression or activity to drive heterotic growth. In this work, we show that accumulation of growth-promoting and energy metabolism proteins, enhanced energy metabolism activities, and increased protein lysine acetylation were associated with superior growth of the panicle meristem in the elite hybrid rice Shanyou 63 relative to its parental varieties. Metabolism of nuclear/cytosolic acetyl-coenzyme A was also enhanced in the hybrid, which paralleled increases in histone H3 acetylation to selectively target the expression of growth-promoting and metabolic genes. Lysine acetylation of cellular proteins, including TARGET OF RAPAMYCIN complex 1, ribosomal proteins, and energy metabolism enzymes, was also augmented and/or remodeled to modulate their activities. The data indicate that an enhanced network of energy-producing metabolic activity and growth-promoting histone acetylation/gene expression in the hybrid could contribute to its superior growth rate and may constitute a model to explain heterosis.

New Dielis species and structural dichotomy of the mitochondrial cox2 gene in Scoliidae wasps

Some mitochondrial protein-coding genes of protists and land plants have split over the course of evolution into complementary genes whose products can form heteromeric complexes that likely substitute for the undivided proteins. One of these genes, cox2, has also been found to have split in animals, specifically in Scoliidae wasps (Hymenoptera: Apocrita) of the genus Dielis (Campsomerini), while maintaining the conventional structure in related Scolia (Scoliini). Here, a hitherto unrecognized Nearctic species of Dielis, D. tejensis, is described based on its phenotype and mtDNA. The mitogenome of D. tejensis sp. nov. differs from that of the sympatric sibling species Dielis plumipes fossulana by the reduced size of the cox2-dividing insert, which, however, still constitutes the fifth part of the mtDNA; an enlarged nad2-trnW intergenic region; the presence of two trnKttt paralogues; and other features. Both species of Dielis have a unique insertion of a threonine in COXIIA, predicted to be involved in COXIIA-COXIIB docking, and substitutions of two hydrophobic residues with redox-active cysteines around the CuA centre in COXIIB. Importantly, the analysis of mtDNA from another Campsomerini genus, Megacampsomeris, shows that its cox2 gene is also split. The presented data highlight evolutionary processes taking place in hymenopteran mitogenomes that do not fall within the mainstream of animal mitochondrion evolution.

Genetics for seed traits and Mungbean yellow mosaic India virus reaction in urdbean (Vigna mungo L. Hepper)

AbstractUrdbean or blackgram is one of the important multi‐season pulse crops grown in India. Improved seed quality and resistance to Mungbean yellow mosaic India virus (MYMIV) are the important objectives of the urdbean breeding program. For this, knowledge of inheritance for seed quality traits and MYMIV resistance is essentially required. Therefore, an attempt has been made to understand the inheritance of seed traits like seed luster, seed coat color, mosaic on seed surface, and MYMIV reaction using intra‐ and inter‐specific bi‐parental populations. The inheritance results revealed that in DPU88‐31 × LBG‐685 population, the shining seed registered dominance over the dull trait, while the green seed coat color exhibited dominance over the brown seed coat color. However, the inter‐specific F2 population derived from IPU11‐02 × Pant M 6 deviated significantly from Mendelian ratios and supported severe segregation distortion for seed traits, while the mosaic on seed coat or mottling character studied in interspecific population demonstrated a dominant digenic inheritance pattern. In five inter‐ and intra‐specific F2 populations, MYMIV resistance demonstrated monogenic dominant inheritance, which was also validated in F3 generations. Furthermore, linkage analysis exhibited an association between seed luster and seed coat color, while no association was noticed between seed traits and MYMIV resistance. The normality tests revealed that seed width and 100 seeds weight were controlled by a few major genes, while other quantitative traits were governed by many genes with small additive effects. The skewness suggested that complementary gene interactions were present among genes controlling 100 seed weight in the intra‐specific cross, while duplicate gene interactions were involved in the inter‐specific cross. The identified monogenic traits and linked morphological marker after further enrichment of the linkage group could be used as an important tool in the regular breeding program.

Assessment of frequency distribution for the morphological and yield related traits in BC1F1, BC2F1 and BC2F2 populations in sweet corn

The nature of gene action and the number of genes controlling the traits are the vital criteria for the selection of traits. The present study is to analyze the range of variations within the population and as well as the skewness and kurtosis in backcross and selfed sweet corn populations USC1-2-3-1× UMI1230β+ and SC1107×UMI1230β+. In the BC2F2 population of SC1107×UMI1230β+, most of the traits showed positive skewness and platykurtic distribution which indicated that complementary gene action&nbsp; played a role and more number of genes is controlling the traits. In USC1-2-3-1× UMI1230β+, single plant yield recorded positive skewness. In some traits such as, the number of tassel branches, cob girth, and leaf breadth showed negative skewness and platykurtic distribution was also observed in both the cross combination which indicated prevalence of duplicate gene action. In order to fix an individual trait and for rapid genetic gain, duplicate gene action is required. &nbsp;Keywords: Sweet corn, Skewness, Kurtosis, Gene action.&nbsp

Variability, Correlation and Path Analysis for Several Quantitative Traits Derived Multi-Parent Advanced Generation Inter-Cross (Magic) F2 Population of Rice (Oryza sativa L.)

Eight quantitative traits of an F2 MAGIC population in rice were subjected to analyze variability, correlation and path analysis. The results of third and fourth degree statistics (skewness and kurtosis) showed that Individual dried straw weight, 1000-seed weight, Individual grain yield, and Number of effective tillers are associated with complementary gene interaction, requiring intense selection, while Plant height, Panicle length, Spikelet fertility percentage, and Harvest index are associated with duplicate (additive x additive) gene interactions, are suitable for mild selection. Leptokurtic distribution of Plant height, Number of effective tillers per plant, Pancile length and Spikelet fertility percentage revealed that the large number of genes in governing the traits. Platykurtic distribution of Individual dried straw weight, 1000-seed weight, Individual grain yield and Harvest index indicated fewer genes controlling the traits. However, when it comes to H2 and GAM, high values were seen in Number of effective tillers per plant, Individual dried straw weight, Individual grain yield, the traits could be explained by additive gene action and achieved through mass selection. High heritability and lower GAM of Plant height, Panicle length, 1000-seed weight and Harvest index indicated by non-additive genes with limiting the chances of direct selection. For the Pearson correlation, 1000-seed weight showed no correlation with other traits. Individual grain yield highly correlated with Number of effective tillers per plant (0.864**) and Individual dried straw weight (0.828**). The highest total effects on Individual grain yield were Number of effective tillers per plant (2.7437) and Individual dried straw weight (1.0865). Individual dried straw weight had highest direct effect on Individual grain yield at 1.0734, while most of the traits showed high indirect effect on Individual grain yield through Individual dried straw weight.

Inheritance Pattern of Important Qualitative Traits in Safflower (Carthamus tinctorious L.)

Safflower is an important oilseed crop with poorly studied genetic inheritance of the characters. In this study genetically diverse parents were selected which had many contrasting qualitative traits. The parents used in the study were GMU-1217, GMU- 6854, GMU-6891, GMU-2830, EC 755673 and EC755664. Crosses were made between the parents. Observations of parental traits, traits appearing in F1 and F2 were studied and the data were analysed for goodness of fit by χ2 test for appropriate ratio. A total of seven qualitative characters were studied, they were, branching type, mode of branching, mode of spininess, length of bract, leaf dentation, form of leaf shape, flower colour change at maturity, segregation in F2 generation for different traits showed the presence of either complementary or inhibitory gene action for all the characters studied while other crosses were found to have a combination of both monogenic and digenic gene action.A proper knowledge of inheritance of Agronomic traits helps in planning of efficient strategy for further crop improvement.

Engineered celastrol and plasmid co-delivery for in situ expression and targeted mitochondrial relocation of Nur77 protein towards effective drug resistance reversion

Overexpression of Bcl-2 (B-cell lymphoma-2) is one of the important reasons for the drug resistance of tumor cells, which often leads to the failure of clinical cancer treatment. In recent years, studies have shown that the nuclear orphan receptor Nur77 can interact with Bcl-2 to change the Bcl-2 protein from an anti-apoptotic conformation to a pro-apoptotic conformation, thereby inducing apoptosis. However, since Nur77 needs to migrate from the nucleus to the mitochondria to reverse the anti-apoptotic effect of Bcl-2, it is difficult to effectively activate the Nur77-Bcl-2 apoptotic pathway by the existing system. Herein, we design an intelligent drug delivery system (PPP/Nur77/Cela) in which drug molecular-guided gene expression products actively act on Bcl-2 proteins, which consists of a mPEG-PCL-PEI (methoxy poly(ethylene glycol)-poly(ε-caprolactone)-poly(ethylene imine), or PPP) polymer loaded with Nur77 gene and celastrol (Cela) via electrostatic and hydrophobic interactions. The system can effectively improve solubility and toxicity of Cela, and can express Nur77 efficiently in cells with a gene transfection efficiency comparable to that of PEI. Interestingly, besides being an anticancer drug, Cela can also promote Nur77 to transfer to mitochondria and convert Bcl-2 from anti-apoptotic protein to pro-apoptotic protein, thereby increasing the sensitivity of Bcl-2 overexpression cancer cells to drugs, which achieves efficient cancer treatment through complementary synergistic chemotherapy and gene therapy. To the best of our knowledge, this is a pioneering strategy to use drug molecules to guide gene expression products to actively act on target proteins, which is a truly successful realization of complementary intensive chemotherapy and gene therapy in the treatment of drug-resistant tumors.

Genetic characterization and comparative assessment of response of biparental and multi-parental populations to northern corn leaf blight in maize (Zea mays L.)

To compare the efficiency of multiparental and biparental progenies to northern corn leaf blight (NCLB) resistance, F3 progenies were evaluated during two seasons. A total of 418 F3 progenies derived from multiple founder parents and 248 F3 progenies derived from two parents, namely, SKV50 (Resistant) and CM202 (Susceptible) were phenotyped for their response to the NCLB during summer and kharif 2020. A wide range of disease severity was observed among F3 progenies derived from multiple parents ranging from 28.12 to 62.93%. The difference in the variances between multiparent and biparental progenies was indicated by the significance of Levene’s test. High genotypic and phenotypic coefficient of variation was observed among multi-parental progenies than those derived from the biparental population. Reaction to NCLB showed moderate heritability of 57% and high genetic advance as per cent mean (20.70%) in the case of biparental population, whereas, multi-parental population exhibited a high heritability (68%) and high genetic advance as per cent mean (28.57%). The distribution of F3 progenies was found to be positively skewed and platykurtic indicating that the trait was under the control of large number of genes with complementary gene action.

Genetic variation of morphological and yield-related traits in backcrossed and selfed population of maize

The number of genes controlling the morphological and yield-related traits and their mode of action plays a vital role in selecting traits and favourable progenies in a segregating population. In this study, we analysed the skewness and kurtosis in the backcrossed and selfed populations of SC11-2×UMI1230β1+. Negative skewness refers to redundant gene activity, positive skewness is connected to complementary gene action. Positive kurtosis suggests the existence of gene activity controlled by a smaller number of genes. Negative kurtosis shows a lack of gene activity and is controlled by a large number of genes. The BC2F2 population of plant height and single plant yield showed positive skewness and platykurtic distribution, suggesting that the genes were governed by complementary gene action and controlled by the large number of genes. This implied need for rigorous selection to achieve a genetic gain in later generations. Cob length and kernel weight showed a duplicate gene action which implied that mild selection can fix these traits and accelerate genetic gain.

Low Dopamine D2 Receptor Expression Drives Gene Networks Related to GABA, cAMP, Growth and Neuroinflammation in Striatal Indirect Pathway Neurons

BackgroundA salient effect of addictive drugs is to hijack the dopamine reward system, an evolutionarily conserved driver of goal-directed behavior and learning. Reduced dopamine type 2 receptor availability in the striatum is an important pathophysiological mechanism for addiction that is both consequential and causal for other molecular, cellular, and neuronal network differences etiologic for this disorder. Here, we sought to identify gene expression changes attributable to innate low expression of the Drd2 gene in the striatum and specific to striatal indirect medium spiny neurons (iMSNs). MethodsCre-conditional, translating ribosome affinity purification (TRAP) was used to purify and analyze the translatome (ribosome-bound messenger RNA) of iMSNs from mice with low/heterozygous or wild-type Drd2 expression in iMSNs. Complementary electrophysiological recordings and gene expression analysis of postmortem brain tissue from human cocaine users were performed. ResultsInnate low expression of Drd2 in iMSNs led to differential expression of genes involved in GABA (gamma-aminobutyric acid) and cAMP (cyclic adenosine monophosphate) signaling, neural growth, lipid metabolism, neural excitability, and inflammation. Creb1 was identified as a likely upstream regulator, among others. In human brain, expression of FXYD2, a modulatory subunit of the Na/K pump, was negatively correlated with DRD2 messenger RNA expression. In iMSN-TRAP-Drd2HET mice, increased Cartpt and reduced S100a10 (p11) expression recapitulated previous observations in cocaine paradigms. Electrophysiology experiments supported a higher GABA tone in iMSN-Drd2HET mice. ConclusionsThis study provides strong molecular evidence that, in addiction, inhibition by the indirect pathway is constitutively enhanced through neural growth and increased GABA signaling.

P8.038: Prevalence of Genetic Mutations Associated With Kidney Disorders in Renal Transplant Recipients – Single Centre Experience

Introduction: End-stage renal disease (ESRD) of undetermined etiology is highly prevalent and constitutes a significant clinical challenge, particularly in the context of kidney transplantation. Underlying renal conditions leading to ESRD are extremely heterogeneous and the prevalence of hereditary nephropathies among adults with ESRD is less known. To date, less than 10% of adult ESRD is thought to be genetic. Genetic analysis often reveals the cause of kidney failure in patients with unknown kidney disease. For patients awaiting renal transplantation, genetic testing prior to kidney transplant may help to understand the outcome in these patients. We studied the prevalence of genetic mutations related to kidney disease in renal transplant (RTx) recipients. Methods: It was a Single Centre, Observational, Cohort Study. Between October 2019 & July 2021, 123 RTx recipients were analyzed using Multiplex Ligation-dependent Probe Amplification (MLPA) and clinical exome sequencing for mutations related to kidney disease. Results: Mean age was 36±12 years, 96 (78%) were Males. 88 (71.5%) were undetermined and 35 (28.4 %) had established cause of CKD. 8 patients had structural or hereditary kidney disease and 27 were non-hereditary including DKD, Glomerulopathy, Obstructive uropathy and CTID diagnosed by kidney biopsy and clinical and radiological presentations. 17/88 (19.3%) patients with undetermined CKD on renal biopsy showed Glomerulosclerosis (6), CGN (8) & TMA (2). one had advanced renal damage. 32/88 (36.36%) patients had genetic mutations related to kidney disease. 20 were structural congenital or hereditary kidney disease & 10 had a mutation for Glomerulopathies. Mutations related to complement regulatory genes were found in 84/123 (68.2%). 6 patients had congenital or hereditary kidney disease, 12 were DKD, 5 were Glomerulopathy, 2 had Obstructive Kidney disease and 1 had CTID. Homozygousity and heterozygosity were seen in 14 (16.8%) and 10 (12%) patients respectively. 67/88 (80.7%) also showed CFHR1/CFHR3 gene complex duplication, reported as a variant of uncertain significance. 1 patient had a mutation for the PLG gene, previously been implicated in the pathogenesis of ahus. Conclusion: This study demonstrates 82.9% (102/123) prevalence of the genetic mutation in RTx recipients. Genetic analysis provides the etiology in 35 patients with undetermined kidney disease. The significance of complementary regulatory gene related mutation remained uncertain.

Potato virus X-delivered CRISPR activation programs lead to strong endogenous gene induction and transient metabolic reprogramming in Nicotiana benthamiana.

SUMMARYProgrammable transcriptional regulators based on CRISPR architecture are promising tools for the induction of plant gene expression. In plants, CRISPR gene activation is effective with respect to modulating development processes, such as the flowering time or customizing biochemical composition. The most widely used method for delivering CRISPR components into the plant is Agrobacterium tumefaciens‐mediated genetic transformation, either transient or stable. However, as a result of their versatility and their ability to move, virus‐derived systems have emerged as an interesting alternative for supplying the CRISPR components to the plant, in particular guide RNA (gRNA), which represents the variable component in CRISPR strategies. In the present study, we describe a Potato virus X‐derived vector that, upon agroinfection in Nicotiana benthamiana, serves as a vehicle for delivery of gRNAs, producing highly specific virus‐induced gene activation. The system works in combination with a N. benthamiana transgenic line carrying the remaining complementary CRISPR gene activation components, specifically the dCasEV2.1 cassette, which has been shown previously to mediate strong programmable transcriptional activation in plants. Using an easily scalable, non‐invasive spraying method, we show that gRNA‐mediated activation programs move locally and systemically, generating a strong activation response in different target genes. Furthermore, by activating three different endogenous MYB transcription factors, we demonstrate that this Potato virus X‐based virus‐induced gene reprogramming strategy results in program‐specific metabolic fingerprints in N. benthamiana leaves characterized by distinctive phenylpropanoid‐enriched metabolite profiles.

Investigating the Role of the NLRP3 Inflammasome Pathway in Acute Intestinal Inflammation: Use of THP-1 Knockout Cell Lines in an Advanced Triple Culture Model.

The NLRP3 inflammasome plays an important role in intestinal homeostasis as well as inflammation. However, in vivo studies investigating the role of the NLRP3 inflammasome in inflammatory bowel disease (IBD) report contrasting results, leaving it unclear if the NLRP3 inflammasome augments or attenuates intestinal inflammation. To investigate the role of the NLRP3/caspase-1 pathway in a model of acute intestinal inflammation, we modified a previously established in vitro triple culture model of the healthy and inflamed intestine (Caco-2/HT29-MTX-E12/THP-1). Using THP-1 knockout cell lines, we analyzed how the NLRP3 inflammasome and its downstream enzyme caspase-1 (CASP1) affect inflammatory parameters including barrier integrity and cytotoxicity, as well as gene expression and secretion of pro-inflammatory cytokines and mucus. Furthermore, we investigated differences in inflammation-mediated cytotoxicity towards enterocyte-like (Caco-2) or goblet-like (HT29-MTX-E12) epithelial cells. As a complementary approach, inflammation-related cytotoxicity and gene expression of cytokines was analyzed in intestinal tissue explants from wildtype (WT) and Nlrp3-/- mice. Induction of intestinal inflammation impaired the barrier, caused cytotoxicity, and altered gene expression of pro-inflammatory cytokines and mucins in vitro, while the knockout of NLRP3 and CASP1 in THP 1 cells led to attenuation of these inflammatory parameters. The knockout of CASP1 tended to show a slightly stronger attenuating effect compared to the NLRP3 knockout model. We also found that the inflammation-mediated death of goblet-like cells is NLRP3/caspase-1 dependent. Furthermore, inflammation-related cytotoxicity and upregulation of pro-inflammatory cytokines was present in ileal tissue explants from WT, but not Nlrp3-/- mice. The here presented observations indicate a pro-inflammatory and adverse role of the NLRP3 inflammasome in macrophages during acute intestinal inflammation.

The telomere-to-telomere gap-free genome of four rice parents reveals SV and PAV patterns in hybrid rice breeding.

The telomere-to-telomere gap-free genome of four rice parents reveals SV and PAV patterns in hybrid rice breeding.