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Clustered regularly interspaced short palindromic repeats (CRISPR) is a promising innovative technology for genomic editing that offers scientists the chance to edit DNA structures and change gene function. It has several possible uses consisting of editing inherited deficiencies, treating, and reducing the spread of disorders. Recently, reports have demonstrated the creation of synthetic RNA molecules and supplying them alongside Cas9 into genome of eukaryotes, since distinct specific regions of the genome can be manipulated and targeted. The therapeutic potential of CRISPR/Cas9 technology is great, especially in gene therapy, in which a patient-specific mutation is genetically edited, or in the treating of human disorders that are untreatable with traditional treatments. This review focused on numerous, in vivo , in vitro, and ex vivo uses of the CRISPR/Cas9 technology in human inherited diseases, discovering the capability of this versatile in medicine and examining some of the main limitations for its upcoming use in patients. In addition to introducing a brief impression of the biology of the CRISPR/Cas9 scheme and its mechanisms, we presented the utmost recent progress in the uses of CRISPR/Cas9 technology in editing and treating of human genetic diseases.

Agro-industrial wastes are rich in polyphenols and other bioactive compounds, and valorizing these wastes is a crucial worldwide concern for saving health and the environment. In this work, olive leaf waste was valorized by silver nitrate to produce silver nanoparticles (OLAgNPs), which exhibited various biological, antioxidant, anticancer activities against three cancer cell lines, and antimicrobial activity against multi-drug resistant (MDR) bacteria and fungi. The obtained OLAgNPs were spherical, with an average size of 28 nm, negatively charged at −21 mV, and surrounded by various active groups more than the parent extract based on FTIR spectra. The total phenolic and total flavonoid contents significantly increased in OLAgNPs by 42 and 50% over the olive leaf waste extract (OLWE); consequently, the antioxidant activity of OLAgNPs increased by 12% over OLWE, recording an SC50 of OLAgNPs of 5 µg/mL compared to 30 µg/mL in the extract. The phenolic compound profile detected by HPLC showed that gallic acid, chlorogenic acid, rutin, naringenin, catechin, and propyl gallate were the main compounds in the HPLC profile of OLAgNPs and OLWE; the content of these compounds was higher in OLAgNPs than OLWE by 16-fold. The higher phenolic compounds in OLAgNPs are attributable to the significant increase in biological activities of OLAgNPs than that of OLWE. OLAgNPs successfully inhibited the proliferation of three cancer cell lines, MCF-7, HeLa, and HT-29, by 79–82% compared to 55–67% in OLWE and 75–79% in doxorubicin (DOX). The preliminary worldwide problem is multi-drug resistant microorganisms (MDR) because of the random use of antibiotics. Therefore, in this study, we may find the solution in OLAgNPs with concentrations of 2.5–20 µg/mL, which significantly inhibited the growth of six MDR bacteria L. monocytogenes, B. cereus, S. aureus, Y. enterocolitica, C. jejuni, and E. coli with inhibition zone diameters of 25–37 mm and six pathogenic fungi in the range of 26–35 mm compared to antibiotics. OLAgNPs in this study may be applied safely in new medicine to mitigate free radicals, cancer, and MDR pathogens.

Developing high-yielding and resilient maize hybrids is essential to ensure its sustainable production with the ongoing challenges of considerable shifts in global climate. This study aimed to explore genetic diversity among exotic and local maize inbred lines, evaluate their combining ability, understand the genetic mechanisms influencing ear characteristics and grain yield, and identify superior hybrids suited for timely and late sowing conditions. Seven local and exotic maize inbred lines were genotyped using SSR (Simple Sequence Repeat) markers to assess their genetic diversity. These diverse lines were utilized to develop 21 F1 hybrids using a diallel mating design. These hybrids, alongside a high-yielding commercial check (SC-10), were evaluated under field conditions across two growing seasons under timely and late sowing conditions. The results showed that sowing date, assessed hybrids, and their interaction significantly influenced all measured agronomic traits. Notably, late sowing reduced plant height, ear characteristics, and, ultimately, grain yield. Several hybrids, particularly L101 × L103, L101 × L105, L104 × L105, and L104 × L107 under timely sowing, and L101 × L105 and L104 × L107 under late sowing, surpassed the agronomic performance of check commercial hybrid. Inbred lines L101 and L103 emerged as superior combiners for ear traits and yield, while line L106 showed promise for breeding shorter-stature plants. Hybrid combinations L101 × L105, L104 × L107, and L106 × L107 were identified as specific good combiners for grain yield and related traits under both sowing conditions, indicating their potential for commercial development. Strong positive associations were observed between grain yield and certain agronomic traits highlighting their utility for indirect selection in early breeding generations.

DNA topoisomerase 1 alpha (TOP1α) plays a critical role in plant development, yet its specific involvement in male reproduction is not well understood. Here, we show that TOP1α is essential for Arabidopsis anther development, and its absence leads to severe morphological and functional defects. The top1α1 mutant displays pleiotropic phenotypes, including early flowering and twisted petals, most notably the loss of two microsporangia. This loss stems from disrupted cell division in the L2 layer. Cytological analyses reveal aberrant pollen wall formation and reduced fertility in the mutant. RNA-seq data from top1α1 mutants identify over 2,000 differentially expressed genes, with significant downregulation of key anther development regulators such as SPL , AMS , and MS1 , alongside genes involved in proteostasis and lipid metabolism. Furthermore, chromatin immunoprecipitation (ChIP) confirms TOP1α directly binds to the promoters of crucial genes like AG , MS1 , and MYB80 , suggesting a role in modulating transcriptional accessibility. All observed defects were rescued by complementation with gTOP1α-4HA, confirming the specificity of the top1α1 phenotype. Our findings establish TOP1α as a central coordinator of anther morphogenesis, functioning through intricate gene regulatory networks and chromatin remodeling. This work provides novel insights into the molecular mechanisms governing male reproductive development in plants.

Salinity in soil or irrigation water requires developing genetically salt-tolerant genotypes, especially in arid regions. Developing salt-tolerant and high-yielding wheat genotypes has become more urgent in particular with continuing global population growth and abrupt climate changes. The current study aimed at investigating the genetic variability of new breeding lines in three advanced generations F6–F8 under salinity stress. The evaluated advanced lines were derived through accurate pedigree selection under actual saline field conditions (7.74 dS/m) and using saline water in irrigation (8.35 dS/m). Ninety-four F6 lines were evaluated in 2017–2018 and reduced by selection to thirty-seven F7 lines in 2018–2019 and afterward to thirty-four F8 lines in 2019–2020 based on grain yield and related traits compared with adopted check cultivars. Significant genetic variability was detected for all evaluated agronomic traits across generations in the salt-stressed field. The elite F8 breeding lines displayed higher performance than the adopted check cultivars. These lines were classified based on yield index into four groups using hierarchical clustering ranging from highly salt-tolerant to slightly salt-tolerant genotypes, which efficiently enhance the narrow genetic pool of salt-tolerance. The detected response to selection and high to intermediate broad-sense heritability for measured traits displayed their potentiality to be utilized through advanced generations under salinity stress for identifying salt-tolerant breeding lines.

Wheat, a staple cereal crop, faces challenges due to climate change and increasing global population. Maintaining genetic diversity is vital for developing drought-tolerant cultivars. This study evaluated the genetic diversity and drought response of five wheat cultivars and their corresponding F1 hybrids under well-watered and drought stress conditions. Molecular profiling using ISSR and SCoT-PCR markers revealed 28 polymorphic loci out of 76 amplified. A statistically significant impact of parental genotypes and their crosses was observed on all investigated agro-morphological traits, including root length, root weight, shoot length, shoot weight, proline content, spikelet number/spike, spike length, grain number/spike, and grain weight/spike. The parental genotypes P1 and P3 had desirable positive and significant general combining ability (GCA) effects for shoot fresh weight, shoot dry weight, root fresh weight, root dry weight, shoot length, and root length under well-watered conditions, while P3 and P5 recorded the highest GCA estimates under drought stress. P3 and P4 showed the highest GCA effects for number of spikelets per spike, the number of grains per spike, and grain weight per spike under normal conditions. P5 presented the maximum GCA effects and proved to be the best combiner under drought stress conditions. The cross P1× P3 showed the highest positive specific combining ability (SCA) effects for shoot fresh weight under normal conditions, while P2×P3 excelled under water deficit conditions. P1× P2, P1 × P3, and P4× P5 were most effective for shoot dry weight under normal conditions, whereas P1×P3 and P3×P5 showed significant SCA effects under drought stress. Positive SCA effects for root fresh weight and shoot length were observed for P3×P5 under stressed conditions. Additionally, P4×P5 consistently recorded the highest SCA for root length in both environments, and P3×P5 excelled in the number of spikelets, grains per spike, and grain weight per spike under drought conditions. The evaluated genotypes were categorized based on their agronomic performance under drought stress into distinct groups ranging from drought-tolerant genotypes (group A) to drought-sensitive ones (group C). The genotypes P5, P2×P5, and P3×P5 were identified as promising genotypes to improve agronomic performance under water deficit conditions. The results demonstrated genetic variations for drought tolerance and highlighted the potential of ISSR and SCoT markers in wheat breeding programs for developing drought-tolerant cultivars.

Determining and improving drought-tolerant cultivars is a major goal of plant breeding to face climate change. The productivity of faba bean in Egypt is affected by abiotic stresses, especially drought stress. This study evaluated eight Egyptian faba bean cultivars for drought tolerance under three soil water regimes consisting of well-watered (100% field capacity), moderate drought stress (50% field capacity), and severe drought stress (25% field capacity) regimes in pots under greenhouse conditions using biochemical, physiochemical, and molecular parameters. The cultivars Nubariya 1, Nubariya 3, and Giza 716 showed the highest proline content values under 50% field capacity conditions, with 4.94, 4.39, and 4.26 mmol/g fresh weights, respectively. On the other hand, the cultivars Sakha 1, Sakha 4, Nubariya 1, and Nubariya 3 exhibited the highest proline contents (7.8, 7.53, 6.17, and 6.25, respectively) under 25% field capacity treatment. The molecular profiling was conducted using SCoT and SRAP approaches. Fresh leaves were utilized to extract the DNA, and ten primers for SRAP and six for SCoT were used in the PCR procedures. SCoT and SRAP-PCR generated 72 loci, of which, 55 were polymorphic, and 17 were monomorphic. SCoT and SRAP each had 48 and 24 total loci, respectively. The average polymorphism (%) values achieved via SCoT and SRAP were 70.93% and 80%, respectively. Based on the molecular profiles, the cluster analysis identified three clusters. The first cluster comprised Giza 716 cultivars; the second cluster included Sakha 1, Sakha 3, Sakha 4, and Akba 3300 cultivars; the third cluster comprised two cultivars Nubariya 1 and Nubariya3. According to the study’s findings, Sakha 1, Sakha 4, Nubariya 1, and Nubariya 3 are remarkable parents for developing drought-tolerant faba bean genotypes. Additionally, this study concluded that SRAP and SCoT markers recreated trustworthy banding profiles to evaluate the genetic polymorphism among faba bean cultivars, which are regarded as the cornerstone for genetic improvements in crops.

The gluten strength and the composition of high- and low-molecular-weight glutenin subunits (HMWGSs and LMWGSs) of fifty-one durum wheat genotypes were evaluated using sodium dodecyl sulfate (SDS) sedimentation testing and SDS polyacrylamide gel electrophoresis (SDS-PAGE). This study examined the allelic variability and the composition of HMWGSs and LMWGSs in T. durum wheat genotypes. SDS-PAGE was proven to be a successful method for identifying HMWGS and LMWGS alleles and their importance in determining the dough quality. The evaluated durum wheat genotypes with HMWGS alleles 7+8, 7+9, 13+16, and 17+18 were highly correlated with improved dough strength. The genotypes containing the LMW-2 allele displayed stronger gluten than those with the LMW-1 allele. The comparative in silico analysis indicated that Glu-A1, Glu-B1, and Glu-B3 possessed a typical primary structure. The study also revealed that the lower content of glutamine, proline, glycine, and tyrosineand the higher content of serine and valine in the Glu-A1 and Glu-B1 glutenin subunits, and the higher cysteine residues in Glu-B1 and lower arginine, isoleucine, and leucine in the Glu-B3 glutenin, are associated with the suitability of durum wheat for pasta making and the suitability of bread wheat with good bread-making quality. The phylogeny analysis reported that both Glu-B1 and Glu-B3 had a closer evolutionary relationship in bread and durum wheat, while the Glu-A1 was highly distinct. The results of the current research may help breeders to manage the quality of durum wheat genotypes by exploiting the allelic variation in glutenin. Computational analysis showed the presence of higher proportions of glutamine, glycine, proline, serine, and tyrosine than the other residues in both HMWGSs and LMWGSs. Thus, durum wheat genotype selection according to the presence of a few protein components effectively distinguishes the strongest from the weakest types of gluten.

Nitrogen is an essential element for maize growth, but excessive application can lead to various environmental and ecological issues, including water pollution, air pollution, greenhouse gas emissions, and biodiversity loss. Hence, developing maize hybrids resilient to low-N conditions is vital for sustainable agriculture, particularly in nitrogen-deficient soils. Combining ability and genetic relationships among parental lines is crucial for breeding superior hybrids under diverse nitrogen levels. This study aimed to assess the genetic diversity of maize inbred lines using simple sequence repeat (SSR) markers and evaluate their combining ability to identify superior hybrids under low-N and recommended conditions. Local and exotic inbred lines were genotyped using SSR markers, revealing substantial genetic variation with high gene diversity (He = 0.60), moderate polymorphism information content (PIC = 0.54), and an average of 3.64 alleles per locus. Twenty-one F1 hybrids were generated through a diallel mating design using these diverse lines. These hybrids and a high yielding commercial check (SC-131) were field-tested under low-N and recommended N conditions. Significant variations (p < 0.01) were observed among nitrogen levels, hybrids, and their interaction for all recorded traits. Additive genetic variances predominated over non-additive genetic variances for grain yield and most traits. Inbred IL3 emerged as an effective combiner for developing early maturing genotypes with lower ear placement. Additionally, inbreds IL1, IL2, and IL3 showed promise as superior combiners for enhancing grain yield and related traits under both low-N and recommended conditions. Notably, hybrids IL1×IL4, IL2×IL5, IL2×IL6, and IL5×IL7 exhibited specific combining abilities for increasing grain yield and associated traits under low-N stress conditions. Furthermore, strong positive associations were identified between grain yield and specific traits like plant height, ear length, number of rows per ear, and number of kernels per row. Due to their straightforward measurability, these relationships underscore the potential of using these traits as proxies for indirect selection in early breeding generations, particularly under low-N stress. This research contributes to breeding nitrogen-efficient maize hybrids and advances our understanding of the genetic foundations for tolerance to nitrogen limitations.