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Plants generate organs continuously during post-embryonic development, thus how to preserve stem cells in changing environments is crucial for their survival. Genotoxic stress threatens genome stability in all somatic cells, whereas, in the meristem, only stem cells undergo cell death in response to DNA damage, followed by stem cell replenishment. It was previously shown that inhibition of downward auxin flow in roots participates in DNA damage-induced stem cell death; however, how cell death is confined to stem cells in tissues with reduced auxin content remains elusive. Here we show that, in response to DNA double-strand breaks, the Aux/IAA family members IAA5 and IAA29, which encode the negative regulators of auxin signaling, were locally induced in vascular stem cells and their daughters in Arabidopsis roots. This is an active response to DNA damage in which the transcription factor SUPPRESSOR OF GAMMA RESPONSE 1 directly induces their expression. In the iaa5 iaa29 double mutant, DNA damage-induced stem cell death was significantly suppressed, while it was fully restored by the expression of a stable form of IAA5 in vascular stem cells. Our genetic data revealed that both the suppression of auxin signaling and reduced auxin content are prerequisite for cell death induction, suggesting the central role in maintaining genome integrity in stem cells.One-sentence summaryThe induction of IAA5 and IAA29 promotes stem cell death in Arabidopsis roots under DNA stress.

Saccharomyces cerevisiae cells activate the Rim101 pathway to adapt to alkaline and salt stresses. Upon activation of this pathway, the transcription factor Rim101 undergoes proteolytic activation and regulates the expression of responsive genes. We found Rim101 to be a short-lived protein with a half-life of approximately 15 min. Its rapid turnover was supposedly mediated by the ubiquitin-proteasome system. Excess accumulation of the processed active Rim101 through its overexpression conferred tolerance to both alkaline and salt stresses in yeast cells; in contrast, it had detrimental effects under cadmium stress condition. Cadmium ion inhibited proteolytic activation of Rim101, implying reciprocal interaction between the Rim101 pathway and cadmium stress. Our results demonstrated yeast cells to be equipped with two protective systems to prevent overaccumulation of the processed active Rim101; Rim101 processing is inhibited when Rim101 level is high, and turnover of processed Rim101 is accelerated when it is abundant. Collectively, the results highlighted the flexible aspect of stress response in yeast cell; the cells not only prevent excess activation of one stress-responsive pathway but also facilitate its attenuation to cope with other environmental stresses.

Inorganic CsSnI3 based perovskite crystals are interesting thermoelectric materials, owing to their unusual electronic properties. Here we report the thermoelectric power performance of a solution-coated CsSnI3 thin film from the viewpoint of carrier concentration optimizations. It was found that the carrier concentration can be changed by altering the aging time of the precursor solution. X-ray photoelectron spectroscopy analysis showed that the concentration of metallic Sn4+ increased as the solution aging time increased. This made possible to explore the relationship between carrier concentration and thermoelectric power factor. After controlling Sn4+ concentrations, we report a power factor of 145.10 μW m−1 K−2 , along with electrical conductivity 106 S/cm and Seebeck coefficient of 117 μV/K, measured at room temperature.

Cells with a loss-of-function mutation in a gene (knockout cells) are powerful tools for characterizing the function of such gene product. However, for essential genes, conditional knockout cell lines must be generated. The auxin-inducible degron (AID) technique enables us to conditionally and rapidly deplete a target protein from various eukaryotic cell lines. A combination of CRISPR-/Cas9-based gene editing and AID technique allows us to generate AID-based conditional knockout cell lines. Using these two techniques, we recently proposed a simple and quick way to generate conditional knockout cells for essential genes. In this chapter, we introduce the reader to the experimental procedures to generate these AID-based conditional knockout cell lines.

Cells with a loss-of-function mutation in a gene (knockout cells) are powerful tools for characterizing the function of such gene product. However, for essential genes, conditional knockout cell lines must be generated. The auxin-inducible degron (AID) technique enables us to conditionally and rapidly deplete a target protein from various eukaryotic cell lines. A combination of CRISPR-/Cas9-based gene editing and AID technique allows us to generate AID-based conditional knockout cell lines. Using these two techniques, we recently proposed a simple and quick way to generate conditional knockout cells for essential genes. In this chapter, we introduce the reader to the experimental procedures to generate these AID-based conditional knockout cell lines.

Environment-friendly tin perovskite solar cells (T-PKSCs) are the most suitable alternative candidate for lead-free PKSCs. However, the photovoltaic performance of such T-PKSCs is far below those of lead-based perovskite solar cells due to an energetic mismatch between the perovskite layer and charge transport layers. Herein, it is shown that, by partial substitution of the A-site cation using diethylammonium iodide (DEAI) substitution, deeper energy levels are obtained. At the same time, the trap density is reduced and the grain size is significantly improved. The fabricated solar cell shows much enhanced efficiency from 7.31% to 10.28%, short-circuit current density from 18.68 to 21.69 mA cm−2, open-circuit voltage from 0.59 to 0.67 V, and fill factor from 0.67 to 0.71 after DEAI substitution. Such an efficiency improvement can be explained by matching energy levels at the interfaces between perovskite layer and the charge transport layers. In addition, after 50 days of storage, the modified T-PKSCs demonstrate high stability maintaining 78% of its initial efficiency, whereas the reference device degrades to 68% during 28 days storage.

Despite lead halide perovskite(top)/silicon(bottom) tandem solar cell achieving >29% efficiency, the toxicity of lead limits the adoption of perovskites in electronic devices. To solve this problem, tin perovskite has been suggested as an alternative candidate to be used as the top cell. However, most of the reports on tin perovskite solar cells are based on narrow band gap materials (band gap: 1.20–1.40 eV), which are not suitable to be used as the tandem top cell. In addition, the efficiency of the previously reported wide band gap lead-free perovskites is not satisfactory, shown to be less than 2.00%. This study explores the fabrication of the wide band gap (1.61 eV) ASnI2Br perovskite solar cells through the optimization of formamidinium and guanidinium content to improve the efficiency from 1.68 to 7.00%. Substituting the A site with larger cation results in a larger crystal size, which translated into higher device stability. Better energy-level alignment within the device significantly enhanced the charge injection and extraction, which translated into higher open-circuit voltage. This work provides proof that tin-halide perovskite solar cells have the potential in the fabrication of lead-free all-perovskite tandem solar cells. This research was supported by JST Mirai Program (JPMJMI17EA) and the Chinese Scholarship Council (CSC202008050034).

In yeast, mitochondria are passed on to daughter cells via the actin cable, motor protein Myo2, and adaptor protein Mmr1. They are released from the actin-myosin machinery after reaching the daughter cells. We report that Mmr1 is rapidly degraded by the ubiquitin-proteasome system in Saccharomyces cerevisiae. Redundant ubiquitin ligases Dma1 and Dma2 are responsible for Mmr1 ubiquitination. Dma1/2-mediated Mmr1 ubiquitination requires phosphorylation, most likely at S414 residue by Ste20 and Cla4. These kinases are mostly localized to the growing bud and nearly absent from mother cells, ensuring phosphorylation and ubiquitination of Mmr1 after the mitochondria enter the growing bud. In dma1Δ dma2Δ cells, transported mitochondria are first stacked at the bud-tip and then pulled back to the bud-neck. Stacked mitochondria in dma1Δ dma2Δ cells exhibit abnormal morphology, elevated respiratory activity, and increased level of reactive oxygen species, along with hypersensitivity to oxidative stresses. Collectively, spatiotemporally-regulated Mmr1 turnover guarantees mitochondrial homeostasis.

Recently, Sn perovskite solar cell (Sn PVK PV) are attracting attention. However, the efficiency was still lower than that of Pb perovskite solar cells. Recently, the Sn PVK PVs with efficiency higher than 10% have been reported from several research groups. The crystal defects include the presence of Sn4+, Sn2+ defect, I- defect, the presence of Sn0, the interstitial I- and so on. In order to decrease these defect densities, we have proposed some processes such as addition of Ge2+ ion, introduction of ethylammonium cation into A site, and surface passivation of perovskite grain boundary with diaminoethane dilute solution. Our results on efficiency enhancement (13%) is explained by the conduction and valence band energy level against carrier trap depth. In addition, an inverted SnPb perovskite solar cells with 23.3% efficiency is discussed from the view point of optimization of energy alignment.

DNA transfection is an important technology in life sciences, wherein nuclear entry of DNA is necessary to express exogenous DNA. Non-viral vectors and their transfection reagents are useful as safe transfection tools. However, they have no effect on the transfection of non-proliferating cells, the reason for which is not well understood. This study elucidates the mechanism through which transfected DNA enters the nucleus for gene expression. To monitor the behavior of transfected DNA, we introduce plasmid bearing lacO repeats and RFP-coding sequences into cells expressing GFP-LacI and observe plasmid behavior and RFP expression in living cells. RFP expression appears only after mitosis. Electron microscopy reveals that plasmids are wrapped with nuclear envelope (NE)‒like membranes or associated with chromosomes at telophase. The depletion of BAF, which is involved in NE reformation, delays plasmid RFP expression. These results suggest that transfected DNA is incorporated into the nucleus during NE reformation at telophase., Haraguchi et al. investigate how transfected DNA is incorporated into the nucleus using light and electron microscopy, and the LacI/LacO system. The authors report that cytoplasmic localised plasmid DNA is incorporated into the nucleus as it reforms during M-exit and transgene expression was achieved only after nuclear reformation in the next G1.

Plant hormones cytokinin and auxin orchestrate differential DNA damage responses in Arabidopsis roots., Plants have a high ability to cope with changing environments and grow continuously throughout life. However, the mechanisms by which plants strike a balance between stress response and organ growth remain elusive. Here, we found that DNA double-strand breaks enhance the accumulation of cytokinin hormones through the DNA damage signaling pathway in the Arabidopsis root tip. Our data showed that activation of cytokinin signaling suppresses the expression of some of the PIN-FORMED genes that encode efflux carriers of another hormone, auxin, thereby decreasing the auxin signals in the root tip and causing cell cycle arrest at G2 phase and stem cell death. Elevated cytokinin signaling also promotes an early transition from cell division to endoreplication in the basal part of the root apex. We propose that plant hormones spatially coordinate differential DNA damage responses, thereby maintaining genome integrity and minimizing cell death to ensure continuous root growth.

In the composition of Q0.1(FA0.75MA0.25)0.9SnI3, Q is replaced with Na+, K+, Cs+, ethylammonium+ (EA+), and butylammonium+ (BA+), respectively, and the relationship between actually measured lattice strain and photovoltaic performances is discussed. The lattice strain evaluated by the Williamson-hall plot of X-ray diffraction data decreased as the tolerance factor was close to one. The efficiency of the Sn-perovskite solar cell was enhanced as the lattice strain decreased. Among them, EA0.1(FA0.75MA0.25)0.9SnI3 having lowest lattice strain gave the best result of 5.41%. Because the carrier mobility increased with a decrease in the lattice strain, these lattice strains would disturb carrier mobility and decrease the solar cell efficiency. Finally, the results that the efficiency of the SnGe-perovskite solar cells was gradually enhanced from 6.42 to 7.60% during storage, was explained by the lattice strain relaxation during the storage.

The efficiency of Sn-perovskite solar (Sn-PVK) cells was enhanced by adding 1% Samarium iodide (SmI2). Increase in Jsc and FF is associated with the enhancement of efficiency and and explained by l...

Although centromeres usually associate with heterochromatic repetitive sequences, such repetitive sequences are not detected around neocentromeres. Nishimura et al. performed systematic 4C analysis of cells containing differently positioned neocentromeres and demonstrate that these neocentromeres commonly associate with distant heterochromatin-rich regions at the 3D level., The centromere is an important genomic locus for chromosomal segregation. Although the centromere is specified by sequence-independent epigenetic mechanisms in most organisms, it is usually composed of highly repetitive sequences, which associate with heterochromatin. We have previously generated various chicken DT40 cell lines containing differently positioned neocentromeres, which do not contain repetitive sequences and do not associate with heterochromatin. In this study, we performed systematic 4C analysis using three cell lines containing differently positioned neocentromeres to identify neocentromere-associated regions at the 3D level. This analysis reveals that these neocentromeres commonly associate with specific heterochromatin-rich regions, which were distantly located from neocentromeres. In addition, we demonstrate that centromeric chromatin adopts a compact structure, and centromere clustering also occurs in vertebrate interphase nuclei. Interestingly, the occurrence of centromere–heterochromatin associations depend on CENP-H, but not CENP-C. Our analyses provide an insight into understanding the 3D architecture of the genome, including the centromeres., Graphical Abstract

The eukaryotic replisome is disassembled in each cell cycle, dependent upon ubiquitylation of the CMG helicase. Studies of Saccharomyces cerevisiae, Caenorhabditis elegans and Xenopus laevis have revealed surprising evolutionary diversity in the ubiquitin ligases that control CMG ubiquitylation, but regulated disassembly of the mammalian replisome has yet to be explored. Here, we describe a model system for studying the ubiquitylation and chromatin extraction of the mammalian CMG replisome, based on mouse embryonic stem cells. We show that the ubiquitin ligase CUL2LRR1 is required for ubiquitylation of the CMG-MCM7 subunit during S-phase, leading to disassembly by the p97 ATPase. Moreover, a second pathway of CMG disassembly is activated during mitosis, dependent upon the TRAIP ubiquitin ligase that is mutated in primordial dwarfism and mis-regulated in various cancers. These findings indicate that replisome disassembly in diverse metazoa is regulated by a conserved pair of ubiquitin ligases, distinct from those present in other eukaryotes.

Because of its small size, Gram-negative Sphingomonas paucimobilis can pose a risk of nosocomial infection. We report the complete circular genome sequence of S. paucimobilis strain Kira, which was isolated from retinoic acid-supplemented SH-SY5Y human cell cultures, to be 3,917,410 bp (G+C content, 65.7%; 3,672 protein-coding sequences), with two plasmids (79,575 bp and 44,333 bp).

The eukaryotic replisome is disassembled in each cell cycle, dependent upon ubiquitylation of the CMG helicase. Studies of Saccharomyces cerevisiae, Caenorhabditis elegans and Xenopus laevis have revealed surprising evolutionary diversity in the ubiquitin ligases that control CMG ubiquitylation, but regulated disassembly of the mammalian replisome has yet to be explored. Here, we describe a model system for studying the ubiquitylation and chromatin extraction of the mammalian CMG replisome, based on mouse embryonic stem cells. We show that the ubiquitin ligase CUL2LRR1 is required for ubiquitylation of the CMG‐MCM7 subunit during S‐phase, leading to disassembly by the p97 ATPase. Moreover, a second pathway of CMG disassembly is activated during mitosis, dependent upon the TRAIP ubiquitin ligase that is mutated in primordial dwarfism and mis‐regulated in various cancers. These findings indicate that replisome disassembly in diverse metazoa is regulated by a conserved pair of ubiquitin ligases, distinct from those present in other eukaryotes., This study reports a model system based on mouse ESC to study the mammalian replisome. It shows that the ubiquitin ligases CUL2LRR1 and TRAIP control p97‐dependent replisome disassembly during DNA replication termination and mitosis, respectively.

Certified efficiency of halogenated Pb-perovskite solar cells has reached 25.2 %. Because of the Pb usage restriction, researches on Pb free perovskite solar cells have been focused on. Halogenated Sn-perovskite is one of candidates for the Pb-free perovskite solar cells. In spite of the similarities of the electronic properties between Pb-perovskite and Sn-perovskite, the efficiency of the Sn-perovskite solar cell was still about 10%, which is far below that of Pb-perovskite solar cells. We report Ge ion doped Sn-perovskite solar cells (Pb free) with 13% efficiency by optimizing A site cations in ASnI3 composition and surface passivation.

Plant root growth is influenced by external factors to adapt to changing environmental conditions. However, the mechanisms by which environmental stresses affect root growth remain elusive. Here we found that DNA double-strand breaks (DSBs) induce the expression of genes for the synthesis of cytokinin hormones and enhance the accumulation of cytokinins in the Arabidopsis root tip. This is a programmed response to DSBs through the DNA damage signaling pathway. Our data showed that activation of cytokinin signalling suppresses the expression of PIN-FORMED genes that encode efflux carriers of another plant hormone, auxin, thereby disturbing downward auxin flow and causing cell cycle retardation in the G2 phase. Elevated cytokinin signalling also promotes an early transition from cell division to endoreplication, resulting in a reduction of the root meristem size. We propose that in response to DNA stress, plants inhibit root growth by orchestrating hormone biosynthesis and signalling.

This is a report on Pb free perovskite solar cells. Direction to enhancing efficiency for Sn-perovskite solar cells are discussed. In order to decrease carrier density and defect density in Sn-perovskite layers, Ge ion doping, surface and grain boundary passivation, decrease in lattice strain, and band alignment are proved to be effective. Ge doped Sn-perovskite solar cells with 13% efficiency is reported.

CENP-A incorporation is critical for centromere specification and is mediated by its chaperone HJURP. The CENP-A-targeting-domain (CATD) of CENP-A specifically binds to HJURP and this binding itself is conserved. However, critical residues of CENP-A vary between species, and the binding interface of CENP-A–HJURP is yet to be understood. Here, we identified the critical residues for chicken CENP-A or HJURP. Interestingly, the A59Q mutation in the α1-helix of chicken CENP-A caused CENP-A mis-incorporation and subsequent cell death, while the corresponding mutation in human CENP-A (S68Q) did not. We also found that W53 of HJURP, which is a contact site of A59 in CENP-A, is also essential in chicken cells. Our comprehensive analyses revealed that the affinity of human HJURP to CATD is strong enough even with the CENP-A α1-helix mutation, but this is not the case in chicken. However, introduction of two arginine residues to the chicken HJURP αA-helix suppressed the CENP-A mis-incorporation in chicken cells expressing CENP-AA59Q. Our data explain the mechanisms and evolution of CENP-A essentiality via CENP-A–HJURP interaction.

Auxin-Inducible Degron (AID) technology enables conditional depletion of targeted proteins. However, the applicability of the AID in vertebrate cells has been limited due to cytotoxicity caused by high auxin concentrations. Here, we establish an improved AID system using an engineered orthogonal auxin-TIR1 pair, which exhibits over 1,000 times stronger binding. With ~1,000-fold less auxin concentration, we achieved to generate the AID-based knockout cells in various human and mouse cell lines in a single transfection.

Tin (Sn) based perovskite solar cells (PSCs) are rapidly getting attention due to their relatively less toxic nature compared to lead-PSCs. However, Sn perovskites previously reported are narrow-bandgap materials. Wide-bandgap perovskites, which are the key materials for the top layer of lead-free tandem solar cells, are rarely researched up to now. Here, it is demonstrated that optoelectronic properties of the GA0.06(FA0.8Cs0.2)0.94SnI2Br based wide-bandgap perovskite can be improved by exploring (1) Germanium (II) Iodide (GeI2) doping, (2) incorporation of Ethylenediamine Bromide (EDABr2) at A site, (3) EDA passivation. The incorporation of big organic cation EDABr2 in the wide-bandgap tin perovskite crystal structure together with GeI2 doping made the carrier lifetime of perovskite absorber increase from 1.1 ns to 22.8 ns and the power conversion efficiency (PCE) was enhanced from 2.55% to 4.86% with an increment of Voc ˃ 100 mV. Further, we achieved the balanced charge transfer by using EDA passivation on the optimized perovskite surface film. As a result, the efficiency was improved from 4.86% to 7.50%, which is the highest efficiency among lead-free wide bandgap PSCs. This enhanced photovoltaic performance of the wide-bandgap tin perovskite device presents a wide application in lead-free tandem towards commercial development.

Generation of cells with a loss-of-function mutation in a gene (knockout cells) is a valuable technique for studying the function of a given gene product. However, if the product of the target gene is essential for cell viability, conditional knockout cell lines must be generated. Recently, as gene editing technology using CRISPR/Cas9 has developed, it has become possible to produce conditional knockout cell lines using this technique. However, to obtain final conditional knockout cell lines, it is necessary to perform several experiments with multiple complicated steps. In this paper, we introduce an easy and efficient method to generate conditional knockout cell lines based on combining auxin-inducible degron (AID) technology with CRISPR/Cas9 gene editing. Our method only requires performing a single transfection and is therefore an easy and rapid method to obtain a conditional knockout cell line.