Plant expression vectors are essential tools for gene functional analysis and molecular plant breeding.The gene of interest is transferred to the vector by molecular cloning technology.Nimble Cloning is a newly develo...Plant expression vectors are essential tools for gene functional analysis and molecular plant breeding.The gene of interest is transferred to the vector by molecular cloning technology.Nimble Cloning is a newly developed molecular cloning method with the advantages of simplicity,efficiency,and standardization.In this study,we developed a"pNC"vector system that contains 55 Nimble Cloning-compatible vectors for functional analysis of genes in plants.These vectors contain the NC frame flanked by unique adapters for one-step and standardized Nimble Cloning.We demonstrate that the pNC vectors are convenient and effective for the functional analysis of plant genes,including the study of gene ectopic expression,protein subcellular localization,protein-protein interaction,gene silencing(RNAi),virus-induced gene silencing,promoter activity,and CRISPR-Cas9-mediated genome editing.The"pNC"vector system represents a high-throughput toolkit that can facilitate the large-scale analysis of plant functional genomics.展开更多
Background:Hypertrophic scarring after burn injury is one of the greatest unmet challenges in patients with burn injuries.A better understanding of the characteristics of scar maturation and early prediction of the lo...Background:Hypertrophic scarring after burn injury is one of the greatest unmet challenges in patients with burn injuries.A better understanding of the characteristics of scar maturation and early prediction of the long-term outcome of scarring are prerequisites for improving targeted therapies and pivotal for patient counselling.Methods:Repeated measurements of scar stiffness in 11 pediatric patients were performed over the course of 1 year using 2 suction devices,the Cutometer and the Nimble.In addition,the observer pliability score of the Patient and Observer Scar Assessment Scale was applied.This longitudinal study allowed quantification of the ability of each of the measured parameters to reflect scar maturation,as indicated by change in skin pliability/stiffness,over time(using linear regression);the ability to distinguish individual patients(intraclass correlation coefficient(ICC));the correlation of the devices(Spearman correlation coefficient);and the ability to predict longterm scar maturation based on early scar assessment(using receiver operating characteristic).Results:All the tools used showed significant longitudinal decrease of scar stiffness from 3 months until 12 months after the injury.The Nimble(ICC_(patient)^(Nimble)=0.99)and the Cutometer(ICC_(patient)^(Cuto)=0.97)demonstrated an excellent ability to distinguish between individual patients.The Nimble seemed to be able to predict the 12-month pliability of scars based on early(3-month)measurements(area under the curve(AUC)_(12m)^(POSAS)=0.67;AUC_(12m)^(C)=0.46;AUC_(12m)^(N)=0.79).Conclusions:The results of this preliminary study suggest that all 3 tools provide suitable means to quantify alterations in scar stiffness over time.Initial evidence suggests the Nimble is most favorable for predicting changes in stiffness associated with long-term scar maturation.Further studies with a larger sample size are required to validate tissue suction as a clinical tool for analysis of changes of scar stiffness over time.展开更多
基金the Hainan Provincial Natural Science Foundation of China(320RC706 and 322RC760)the Central Public-interest Scientific Institution Basal Research Fund(no.1630052022004).
文摘Plant expression vectors are essential tools for gene functional analysis and molecular plant breeding.The gene of interest is transferred to the vector by molecular cloning technology.Nimble Cloning is a newly developed molecular cloning method with the advantages of simplicity,efficiency,and standardization.In this study,we developed a"pNC"vector system that contains 55 Nimble Cloning-compatible vectors for functional analysis of genes in plants.These vectors contain the NC frame flanked by unique adapters for one-step and standardized Nimble Cloning.We demonstrate that the pNC vectors are convenient and effective for the functional analysis of plant genes,including the study of gene ectopic expression,protein subcellular localization,protein-protein interaction,gene silencing(RNAi),virus-induced gene silencing,promoter activity,and CRISPR-Cas9-mediated genome editing.The"pNC"vector system represents a high-throughput toolkit that can facilitate the large-scale analysis of plant functional genomics.
基金supported by grants from the Helmut Horten Foundation。
文摘Background:Hypertrophic scarring after burn injury is one of the greatest unmet challenges in patients with burn injuries.A better understanding of the characteristics of scar maturation and early prediction of the long-term outcome of scarring are prerequisites for improving targeted therapies and pivotal for patient counselling.Methods:Repeated measurements of scar stiffness in 11 pediatric patients were performed over the course of 1 year using 2 suction devices,the Cutometer and the Nimble.In addition,the observer pliability score of the Patient and Observer Scar Assessment Scale was applied.This longitudinal study allowed quantification of the ability of each of the measured parameters to reflect scar maturation,as indicated by change in skin pliability/stiffness,over time(using linear regression);the ability to distinguish individual patients(intraclass correlation coefficient(ICC));the correlation of the devices(Spearman correlation coefficient);and the ability to predict longterm scar maturation based on early scar assessment(using receiver operating characteristic).Results:All the tools used showed significant longitudinal decrease of scar stiffness from 3 months until 12 months after the injury.The Nimble(ICC_(patient)^(Nimble)=0.99)and the Cutometer(ICC_(patient)^(Cuto)=0.97)demonstrated an excellent ability to distinguish between individual patients.The Nimble seemed to be able to predict the 12-month pliability of scars based on early(3-month)measurements(area under the curve(AUC)_(12m)^(POSAS)=0.67;AUC_(12m)^(C)=0.46;AUC_(12m)^(N)=0.79).Conclusions:The results of this preliminary study suggest that all 3 tools provide suitable means to quantify alterations in scar stiffness over time.Initial evidence suggests the Nimble is most favorable for predicting changes in stiffness associated with long-term scar maturation.Further studies with a larger sample size are required to validate tissue suction as a clinical tool for analysis of changes of scar stiffness over time.
