Gene synthesis has provided important contributions in various fields including genomics and medicine. Current genes are 7 - 30 cents depending on the assembly and sequencing methods performed. Demand for gene synthes...Gene synthesis has provided important contributions in various fields including genomics and medicine. Current genes are 7 - 30 cents depending on the assembly and sequencing methods performed. Demand for gene synthesis has been increasing for the past few decades, yet available methods remain expensive. A solution to this problem involves microchip-derived oligonucleotides (oligos), an oligo pool with a substantial number of oligo fragments. Microchips have been proposed as a tool for gene synthesis, but this approach has been criticized for its high error rate during sequencing. This study tests a possible cost-effective method for gene synthesis utilizing fragment assembly and golden gate assembly, which can be employed for quicker manufacturing and efficient execution of genes in the near future. The droplet method was tested in two trials to determine the viability of the method through the accuracy of the oligos sequenced. A preliminary research experiment was performed to determine the efficacy of oligo lengths ranging from two to four overlapping oligos through Gibson assembly. Of the three oligo lengths tested, only two fragment oligos were correctly sequenced. Two fragment oligos were used for the second experiment, which determined the efficacy of the droplet method in reducing gene synthesis cost and speed. The first trial utilized a high-fidelity polymerase and resulted in 3% correctly sequenced oligos, so the second trial utilized a non-high-fidelity polymerase, resulting in 8% correctly sequenced oligos. After calculating, the cost of gene synthesis lowers down to 0.8 cents/base. The final calculated cost of 0.8 cents/base is significantly cheaper than other manufacturing costs of 7 - 30 cents/base. Reducing the cost of gene synthesis provides new insight into the cost-effectiveness of present technologies and protocols and has the potential to benefit the fields of bioengineering and gene therapy.展开更多
Thermo-responsive random copolymers,poly(2-(2-methoxyethoxy)ethoxyethyl methacrylate-co-(ethylene glycol)methyl ether methacrylate)(P(EO_(2)-co-EO_(4/5)))and poly(2-(2-methoxyethoxy)ethoxyethyl methacrylate-co-ethylen...Thermo-responsive random copolymers,poly(2-(2-methoxyethoxy)ethoxyethyl methacrylate-co-(ethylene glycol)methyl ether methacrylate)(P(EO_(2)-co-EO_(4/5)))and poly(2-(2-methoxyethoxy)ethoxyethyl methacrylate-co-ethylene glycol methacrylate(P(EO2-co-EG4/5))are synthesized via atom transfer radical polymerization(ATRP).The successful synthesis and the narrow polydispersity index(PDI)of two copolymers are indicated by 1H nuclear magnetic resonance(1H-NMR)and gel permeation chromatography(GPC)analyses.The transition behaviors of polymers in the aqueous solution are demonstrated by changes in turbidity and particle sizes.The transition behavior of P(EO2-co-EG4/5)is found to be milder than that of P(EO2-co-EO4/5).Moreover,the presence of hydrogen bonds without thermo-responsive properties established by hydroxyl groups in the end-side chain of P(EO_(2)-co-EG_(4/5))hinders the dehydration at the transition temperature(TT).Attenuated total reflection Fourier transform infrared spectrometry(ATR-FTIR)analysis along with contact angle measurements reveals that both P(EO_(2)-co-EO_(4/5))and P(EO_(2)-co-EG_(4/5))films undergo phase transitions from hydrophilicity to hydrophobicity above TT.By examining the swelling and collapse behaviors of the polymer films during phase transitions,it can be concluded that the end hydroxyl groups may establish hydrogen bonds with neighboring ether groups within the films,which remain intact throughout the phase transition process due to their strong bonding interactions.This leads to an increase in steric hindrance within swollen films thereby impeding dehydration processes and inducing hysteresis during phase transitions.展开更多
目的评价巴戟天寡糖胶囊(抗抑郁症中药)治疗抑郁症的疗效与安全性。方法用区组随机、双盲双模拟、安慰剂与阳性药平行对照、多中心临床试验方法,受试者被随机分到3组:巴戟天寡糖胶囊试验组(300 or 600 mg.d-1)、盐酸氟西汀片对照组(20 o...目的评价巴戟天寡糖胶囊(抗抑郁症中药)治疗抑郁症的疗效与安全性。方法用区组随机、双盲双模拟、安慰剂与阳性药平行对照、多中心临床试验方法,受试者被随机分到3组:巴戟天寡糖胶囊试验组(300 or 600 mg.d-1)、盐酸氟西汀片对照组(20 or 30 mg.d-1)及安慰剂组,疗程6周。结果用药6周末,HAMD-17评分与基线差值,巴戟天寡糖胶囊试验组、盐酸氟西汀片对照组、安慰剂组分别为-(13.03±5.43),-(13.86±5.24),-(8.27±4.76),差异有统计学意义(P=0.0000);HAMA与基线差值,3组分别为-(8.87±5.29),-(11.10±5.90),-(6.31±6.07),差异有统计学意义(P=0.0000)。3组有效率分别为71.23%,72.65%,36.97%,差异有统计学意义(P<0.01)。试验组与对照组率差统计学意义(P<0.05)。中医证候疗效,3组有效率分别为72.93%,59.83%,37.82%,试验组与对照组均优于安慰剂组,差异有显著统计学意义(P<0.01)。不良反应发生率3组分别为17.49%,22.76%,21.14%,差异无统计学意义(P>0.05)。结论巴戟天寡糖胶囊治疗抑郁症的疗效优于氟西汀;对抑郁伴发的焦虑也有效;不良反应轻微,安全性好。展开更多
文摘Gene synthesis has provided important contributions in various fields including genomics and medicine. Current genes are 7 - 30 cents depending on the assembly and sequencing methods performed. Demand for gene synthesis has been increasing for the past few decades, yet available methods remain expensive. A solution to this problem involves microchip-derived oligonucleotides (oligos), an oligo pool with a substantial number of oligo fragments. Microchips have been proposed as a tool for gene synthesis, but this approach has been criticized for its high error rate during sequencing. This study tests a possible cost-effective method for gene synthesis utilizing fragment assembly and golden gate assembly, which can be employed for quicker manufacturing and efficient execution of genes in the near future. The droplet method was tested in two trials to determine the viability of the method through the accuracy of the oligos sequenced. A preliminary research experiment was performed to determine the efficacy of oligo lengths ranging from two to four overlapping oligos through Gibson assembly. Of the three oligo lengths tested, only two fragment oligos were correctly sequenced. Two fragment oligos were used for the second experiment, which determined the efficacy of the droplet method in reducing gene synthesis cost and speed. The first trial utilized a high-fidelity polymerase and resulted in 3% correctly sequenced oligos, so the second trial utilized a non-high-fidelity polymerase, resulting in 8% correctly sequenced oligos. After calculating, the cost of gene synthesis lowers down to 0.8 cents/base. The final calculated cost of 0.8 cents/base is significantly cheaper than other manufacturing costs of 7 - 30 cents/base. Reducing the cost of gene synthesis provides new insight into the cost-effectiveness of present technologies and protocols and has the potential to benefit the fields of bioengineering and gene therapy.
基金Fujian External Cooperation project of Natural Science Foundation,China(No.2022I0042)。
文摘Thermo-responsive random copolymers,poly(2-(2-methoxyethoxy)ethoxyethyl methacrylate-co-(ethylene glycol)methyl ether methacrylate)(P(EO_(2)-co-EO_(4/5)))and poly(2-(2-methoxyethoxy)ethoxyethyl methacrylate-co-ethylene glycol methacrylate(P(EO2-co-EG4/5))are synthesized via atom transfer radical polymerization(ATRP).The successful synthesis and the narrow polydispersity index(PDI)of two copolymers are indicated by 1H nuclear magnetic resonance(1H-NMR)and gel permeation chromatography(GPC)analyses.The transition behaviors of polymers in the aqueous solution are demonstrated by changes in turbidity and particle sizes.The transition behavior of P(EO2-co-EG4/5)is found to be milder than that of P(EO2-co-EO4/5).Moreover,the presence of hydrogen bonds without thermo-responsive properties established by hydroxyl groups in the end-side chain of P(EO_(2)-co-EG_(4/5))hinders the dehydration at the transition temperature(TT).Attenuated total reflection Fourier transform infrared spectrometry(ATR-FTIR)analysis along with contact angle measurements reveals that both P(EO_(2)-co-EO_(4/5))and P(EO_(2)-co-EG_(4/5))films undergo phase transitions from hydrophilicity to hydrophobicity above TT.By examining the swelling and collapse behaviors of the polymer films during phase transitions,it can be concluded that the end hydroxyl groups may establish hydrogen bonds with neighboring ether groups within the films,which remain intact throughout the phase transition process due to their strong bonding interactions.This leads to an increase in steric hindrance within swollen films thereby impeding dehydration processes and inducing hysteresis during phase transitions.
文摘目的评价巴戟天寡糖胶囊(抗抑郁症中药)治疗抑郁症的疗效与安全性。方法用区组随机、双盲双模拟、安慰剂与阳性药平行对照、多中心临床试验方法,受试者被随机分到3组:巴戟天寡糖胶囊试验组(300 or 600 mg.d-1)、盐酸氟西汀片对照组(20 or 30 mg.d-1)及安慰剂组,疗程6周。结果用药6周末,HAMD-17评分与基线差值,巴戟天寡糖胶囊试验组、盐酸氟西汀片对照组、安慰剂组分别为-(13.03±5.43),-(13.86±5.24),-(8.27±4.76),差异有统计学意义(P=0.0000);HAMA与基线差值,3组分别为-(8.87±5.29),-(11.10±5.90),-(6.31±6.07),差异有统计学意义(P=0.0000)。3组有效率分别为71.23%,72.65%,36.97%,差异有统计学意义(P<0.01)。试验组与对照组率差统计学意义(P<0.05)。中医证候疗效,3组有效率分别为72.93%,59.83%,37.82%,试验组与对照组均优于安慰剂组,差异有显著统计学意义(P<0.01)。不良反应发生率3组分别为17.49%,22.76%,21.14%,差异无统计学意义(P>0.05)。结论巴戟天寡糖胶囊治疗抑郁症的疗效优于氟西汀;对抑郁伴发的焦虑也有效;不良反应轻微,安全性好。
