Search

The loss of function of the COL1A2 gene can result in osteogenesis imperfecta (OI) types I, II, III, and IV and Ehlers-Danlos syndrome (cardiac valvular and arthrochalasia type).To further investigate the significance of COL1A2 in osteogenesis imperfecta and cardiac valve disease, we created a homozygous COL1A2-/- human embryonic stem cell line (WAe009-A-72) using CRISPR/Cas9. In vivo, the WAe009-A-72 cell line retained typical colony form, a normal karyotype, and robustly expressed pluripotency markers while differentiating into all three germ layers. This cell line is a potential tool for investigating the role of the COL1A2 gene in associated disorders.

Fruit wounding occurred at harvest and transportation requires rapid suberization as a major part of the healing process to prevent infection and desiccation. The focus of this work was to explore the mediation of abscisic acid (ABA) on wound-induced suberization and to determine expression profiles of specific genes involved in wound-induced suberization in tomato fruit. The measurements of weight loss and fruit firmness suggested wound-induced suberization started at 2 d after wounding. The suberization process with the accumulation of suberin polyphenolics (SPP) and polyaliphatics (SPA) observed through autofluorescence microscopy and Sudan IV staining was accelerated by ABA. Expressions of SlPAL5 and Sl4CL involved in the synthesis of SPP reached the highest at 4 and 8 d after wounding following ABA application, respectively. Associated with SPA biosynthesis, SlLACS1 and SlLACS2 showed the most abundant transcripts at 8 and 6 d in ABA group, respectively. Transcript levels including SlKCSs, SlCYP86B1, SlFAR3, and SlGPATs were up-regulated at 2 d after wounding by ABA. Activities of polyphenol oxidase and lipoxygenase were also enhanced during wound-induced suberization following ABA application. The results in this study proved that ABA accelerated the wound-induced suberization progress by increasing the transcript levels of relevant genes in postharvest tomato fruit.

Holt-Oram syndrome (HOS), which is caused by genetic changes in the TBX5 gene, affects the hands and heart. HOS patients have heart defects, including atrial septal defects (ASD), ventricular septal defects (VSD) and heart conduction disease. Here, we generated a homozygous TBX5 knockout human embryonic stem cell (hESC) line (TBX5-KO) using a CRISPR/Cas9 system. The TBX5-KO maintained stem cell like morphology, pluripotency markers, normal karyotype, and could differentiate into all three germ layers in vivo. This cell line can provide an in vitro platform for studying the pathogenic mechanisms and biological function of TBX5 in the heart development.

SNTA1 encodes α1-syntrophin, a scaffold protein, which is a component of the dystrophin-associated protein complex. Additionally, α1-syntrophin interacts with SCN5A and nNOS–PMCA4b complex in cardiomyocytes. SNTA1 is a susceptibility locus for arrhythmia and cardiomyopathy. We generated a homozygous SNTA1 knockout human embryonic stem cell (H9SNTA1KO) using the CRISPR/Cas9 system. H9SNTA1KO maintained pluripotency and a normal karyotype and differentiated into three germ layers in vivo.

Mitogen-activated protein kinase (MPK) cascades consist of a set of kinase types (MPKKKs, MPKKs, MPKs) to establish conserved signal-transducing modules mediating plant growth, development as well as responses to internal and external cues. In this study, the expression patterns of six MPKKK, two MPKK, and 11 MPK genes in wheat in responses to external treatments of phytohormones, including naphthylacetic acid (NAA), abscisic acid (ABA), 6-benzyladenine (6-BA), gibberellin (GA3), salisylic acid (SA), jasmonic acid (JA), and ethylene (ETH), were investigated. Expression analysis revealed that several of the MPK cascade genes are responses to the external phytohormone signaling. Of which, TaMPKKKA;3 is induced by 6-BA and NAA while TaMPK4 repressed by ETH, GA3, SA, and JA; TaMPKKKA, TaMPKKKA;3 and TaMPK1 are down-regulated by ETH and GA3 whereas TaMPK9 and TaMPK12 repressed by ETH and JA in addition that TaMPK12 also repressed by GA3; TaMPK12;1 is down-regulated by ABA, GA3 and SA while TaMPK17 repressed by all exogenous phytonormones examined. TaMPK4, a MPK type gene previously characterized to mediate tolerance to phosphate (Pi) deprivation, was functionally evaluated for its role in mediation of responses of plants to exogenous GA3, ETH, SA, and JA. Results indicated that overexpression and antisense expression of TaMPK4 in tobacco dramatically modify the growth of seedlings upon treatments of GA3, SA and JA, in which the overexpressors behaved deteriorated growth feature whereas the seedlings with antisense expression of TaMPK4 exhibited improved seedling phenotype. The growth behaviors in lines overexpressing or antisensely expressing TaMPK4 are closely associated with the biomass and the corresponding hormone-associated parameters. These results demonstrated that TaMPK4 acts as a critical player in mediating the phytohormone signaling. Our findings have identified the phytohormone-responsive MPK cascade genes in wheat and provided a connection between the phytohormone-mediated responses and the MPK cascade pathways.

Zinc finger protein (ZFP) genes comprise a large and diverse gene family, and are involved in biotic and abiotic stress responses in plants. In this study, a total of 126 ZFP genes classified into various types in wheat were characterized and subjected to expression pattern analysis under inorganic phosphate (Pi) deprivation. The wheat ZFP genes and their corresponding GenBank numbers were obtained from the information of a 4×44K wheat gene expression microarray chip. They were confirmed by sequence similarity analysis and named based on their homologs in Brachypodium distachyon or Oriza sativa. Expression analysis based on the microarray chip revealed that these ZFP genes are categorized into 11 classes according to their gene expression patterns in a 24-h of Pi deprivation regime. Among them, ten genes were differentially up-regulated, ten genes differentially down-regulated, and two genes both differentially up- and down-regulated by Pi deprivation. The differentially up- or down-regulated genes exhibited significantly more or less transcripts at one, two, or all of the checking time points (1, 6, and 24 h) of Pi stress in comparison with those of normal growth, respectively. The both differentially up- and down-regulated genes exhibited contrasting expression patterns, of these, TaWRKY70;5 showed significantly up-regulated at 1 and 6 h and down-regulated at 24 h whereas TaAN1AN20-8;2 displayed significantly upregulated at 1 h and downregulated at 6 h under deprivation Pi condition. Real time PCR analysis confirmed the expression patterns of the differentially expressed genes obtained by the microarray chip. Our results indicate that numerous ZFP genes in wheat respond to Pi deprivation and have provided further insight into the molecular basis that plants respond to Pi deprivation mediated by the ZFP gene family.

Abstract: As a specific type of acid phosphatses, phytases play diverse roles in plants by catalazing the degradation of phytic acid and its derivatives. In this study, a rice phytase gene referred to OsPHY1 has been functionally characterized. OsPHY1 contains a 1 620 bp of open reading frame, encoding a 539-aa polypeptide. A conserve domain metallophosphatase (MPP) (MPP_PAPs), generally harbored in phytase and purple acid phosphatases (PAP), was identified in OsPHY1 (residue 194-398). Phylogenetic analysis revealed that OsPHY1 shares high similarities with phytase genes and PAP-type genes that derived from diverse plant species. The OsPHY1 transcripts were detected to be abundant in germinating seeds, suggesting that this gene plays potential roles on degradation of seed phytic acid and its derivatives during the germination process. Biochemical analysis confirmed that OsPHY1 possesses strong catalytic activities on phytic acid-Na2, with optimal temperature of 57°C and suitable pH of 3.5. Based on transgene analysis, the putative role of OsPHY1 in plants on utilization of phytate was assessed. Under the condition that phytic acid-Na2 was used as sole P source, the OsPHY1-overexpressing tobacco plants behaved higher phytase activities, higher concentrations of Pi, more accumulative amount of total phosphorus, and much more improved growth traits than those of the control plants. Therefore, OsPHY1 is acted as an important component on degradation of the phytins during the seed germination process in rice. Also, OsPHY1 has a potential use on generation of elite crop germplasms with improved use efficiencies on phytate and its derivatives.

The brain barrier is an important structure for metal ion homeostasis. According to studies, lead (Pb) exposure disrupts the transportation of copper (Cu) through the brain barrier, which may cause impairment of the nervous system; however, the specific mechanism is unknown. The previous studies suggested the X-linked inhibitor of apoptosis (XIAP) is a sensor for cellular Cu level which mediate the degradation of the MURR1 domain-containing 1 (COMMD1) protein. XIAP/COMMD1 axis was thought to be an important regulator in Cu metabolism maintenance. In this study, the role of XIAP-regulated COMMD1 protein degradation in Pb-induced Cu disorders in brain barrier cells was investigated. Pb exposure significantly increased Cu levels in both cell types, according to atomic absorption technology testing. Western blotting and reverse transcription PCR (RT-PCR) showed that COMMD1 protein levels were significantly increased, whereas XIAP, ATP7A, and ATP7B protein levels were significantly decreased. However, there were no significant effects at the messenger RNA (mRNA) level (XIAP, ATP7A, and ATP7B). Pb-induced Cu accumulation and ATP7B expression were reduced when COMMD1 was knocked down by transient small interfering RNA (siRNA) transfection. In addition, transient plasmid transfection of XIAP before Pb exposure reduced Pb-induced Cu accumulation, increased COMMD1 protein levels, and decreased ATP7B levels. In conclusion, Pb exposure can reduce XIAP protein expression, increase COMMD1 protein levels, and specifically decrease ATP7B protein levels, resulting in Cu accumulation in brain barrier cells.

Summary: Background: Albuminuria is a marker of vascular dysfunction and is associated with chronic renal and cardiovascular diseases. Data on the association between the longitudinal patterns of weight change early in life and albuminuria later in life are limited. We aimed to identify the body mass index (BMI) trajectory across a 30-year span and evaluate its association with middle-age albuminuria. Methods: Of the 4623 participants aged 6–18-year-old recruited by Hanzhong Adolescent Hypertension Study cohort in northern China from March 10, 1987 to June 3, 2017, a total of 1,825 participants followed up with 6 visits over 30 years were enrolled. Group-based trajectory modeling was used to identify distinct BMI trajectories in longitudinal analyses. Albuminuria was defined as a urinary albumin-to-creatinine ratio (uACR) ≥ 30 mg/g. Findings: Three distinct BMI trajectories were identified: low-increasing (n = 671, 36.8%), moderate-increasing (n = 940, 51.5%), and high-increasing (n = 214, 11.7%); male participants exhibited a steeper increase in BMI than females. The uACR was increased linearly from the low- to high-increasing group. A total of 201 individuals developed albuminuria, with an incidence of 11.0%. Compared with the low-increasing group, the odds ratio (OR) of albuminuria in middle age was 2.13(95% confidence interval [CI]: 1.26 to 3.61) for the high-increasing group after full adjustment for age, sex, smoking, alcohol consumption, marital status, systolic blood pressure, diabetes, and hyperlipidemia. The unadjusted ORs of the high-increasing BMI group were 5.08 (2.76–9.37) for males and 3.45 (1.78–6.69) for females, and the association remained significant in males in the fully adjusted models. Interpretation: Higher BMI trajectories are associated with higher uACR and an increased risk of albuminuria in middle age, especially in males. Identifying long-term BMI trajectories from an early age may assist in predicting the risk of renal diseases and cardiovascular disease later in life. Funding: This work was supported by the National Natural Science Foundation of China (81600327, 82070437, 81870319, 82070549, and 82170437), Natural Science Basic Research Program of Shaanxi Province (2021JM-257 and 2021JM-588), Institutional Foundation of the First Affiliated Hospital of Xi'an Jiaotong University (2019QN-06 and 2021ZXY-14), the Clinical Research Award of the First Affiliated Hospital of Xi'an Jiaotong University of China (XJTU1AF-CRF-2019-004, XJTU1AF2021CRF-021, and XJTU1AFCRF-2017-021), Research Incubation Fund of Xi'an People's Hospital (FZ-61), Grants from the Major Chronic Non-communicable Disease Prevention and Control Research Key Project of the Ministry of Science and Technology of China (2017YFC1307604 and 2016YFC1300104).