Most viruses and transposons serve as effective carriers for the introduction of foreign DNA up to 11 kb into vertebrate genomes.However,their activity markedly diminishes with payloads exceeding 11 kb.Expanding the p...Most viruses and transposons serve as effective carriers for the introduction of foreign DNA up to 11 kb into vertebrate genomes.However,their activity markedly diminishes with payloads exceeding 11 kb.Expanding the payload capacity of transposons could facilitate more sophisticated cargo designs,improving the regulation of expression and minimizing mutagenic risks associated with molecular therapeutics,metabolic engineering,and transgenic animal production.In this study,we improved the Tol2 transposon by increasing protein expression levels using a translational enhancer(QBI SP163,ST)and enhanced the nuclear targeting ability using the nuclear localization protein H2B(SHT).The modified Tol2 and ST transposon efficiently integrated large DNA cargos into human cell cultures(H1299),comparable to the well-established super PiggyBac system.Furthermore,mRNA from ST and SHT showed a significant increase in transgene delivery efficiency of large DNA payloads(8 kb,14 kb,and 24 kb)into zebrafish(Danio rerio).This study presents a modified Tol2 transposon as an enhanced nonviral vector for the delivery of large DNA payloads in transgenic applications.展开更多
Pancreatic ductal adenocarcinoma(PDA)is a devastating disease with pronounced morbidity and a high mortality rate.Currently available treatments lack convincing cost-efficiency determinations and are in most cases not...Pancreatic ductal adenocarcinoma(PDA)is a devastating disease with pronounced morbidity and a high mortality rate.Currently available treatments lack convincing cost-efficiency determinations and are in most cases not associated with relevant success rate.Experimental stimulation of the immune system in murine PDA models has revealed some promising results.Tolllike receptors(TLRs)are pillars of the immune system that have been linked to several forms of malignancy,including lung,breast and colon cancer.In humans,TLRs are expressed in the pancreatic cancer tissue and in several cancer cell lines,whereas they are not expressed in the normal pancreas.In the present review,we explore the current knowledge concerning the role of different TLRs associated to PDA.Even if almost all known TLRs are expressed in the pancreatic cancer microenvironment,there are only five TLRs suggested as possible therapeutic targets.Most data points at TLR2 and TLR9 as effective tumor markers and agonists could potentially be used as e.g.future adjuvant therapies.The elucidation of the role of TLR3 in PDA is only in its initial phase.The inhibition/blockage of TLR4-related pathways has shown some promising effects,but there are still many steps left before TLR4 inhibitors can be considered as possible therapeutic agents.Finally,TLR7 antagonists seem to be potential candidates for therapy.Independent of their potential in immunotherapies,all existing data indicate that TLRs are strongly involved in the pathophysiology and development of PDA.展开更多
基金supported by the National Science and Technology Innovation 2030 Major Projects(2021ZD0202200)National Natural Science Foundation of China(32171090,81970264)+1 种基金Shanghai Science and Technology Commission(21ZR1482600)2023 Youth Innovation Promotion Association CAS。
文摘Most viruses and transposons serve as effective carriers for the introduction of foreign DNA up to 11 kb into vertebrate genomes.However,their activity markedly diminishes with payloads exceeding 11 kb.Expanding the payload capacity of transposons could facilitate more sophisticated cargo designs,improving the regulation of expression and minimizing mutagenic risks associated with molecular therapeutics,metabolic engineering,and transgenic animal production.In this study,we improved the Tol2 transposon by increasing protein expression levels using a translational enhancer(QBI SP163,ST)and enhanced the nuclear targeting ability using the nuclear localization protein H2B(SHT).The modified Tol2 and ST transposon efficiently integrated large DNA cargos into human cell cultures(H1299),comparable to the well-established super PiggyBac system.Furthermore,mRNA from ST and SHT showed a significant increase in transgene delivery efficiency of large DNA payloads(8 kb,14 kb,and 24 kb)into zebrafish(Danio rerio).This study presents a modified Tol2 transposon as an enhanced nonviral vector for the delivery of large DNA payloads in transgenic applications.
基金supported by The National Natural Science Foundation of China (30721064, 30730056, 30620120101)National BasicResearch Program of China (2005CB522504, 2006CB943801,2007CB914502)~~
文摘Pancreatic ductal adenocarcinoma(PDA)is a devastating disease with pronounced morbidity and a high mortality rate.Currently available treatments lack convincing cost-efficiency determinations and are in most cases not associated with relevant success rate.Experimental stimulation of the immune system in murine PDA models has revealed some promising results.Tolllike receptors(TLRs)are pillars of the immune system that have been linked to several forms of malignancy,including lung,breast and colon cancer.In humans,TLRs are expressed in the pancreatic cancer tissue and in several cancer cell lines,whereas they are not expressed in the normal pancreas.In the present review,we explore the current knowledge concerning the role of different TLRs associated to PDA.Even if almost all known TLRs are expressed in the pancreatic cancer microenvironment,there are only five TLRs suggested as possible therapeutic targets.Most data points at TLR2 and TLR9 as effective tumor markers and agonists could potentially be used as e.g.future adjuvant therapies.The elucidation of the role of TLR3 in PDA is only in its initial phase.The inhibition/blockage of TLR4-related pathways has shown some promising effects,but there are still many steps left before TLR4 inhibitors can be considered as possible therapeutic agents.Finally,TLR7 antagonists seem to be potential candidates for therapy.Independent of their potential in immunotherapies,all existing data indicate that TLRs are strongly involved in the pathophysiology and development of PDA.
