The clustered regularly interspaced short palindromic repeats(CRISPR)–CRISPR-associated protein(Cas) system has been widely used for genome editing. In this system, the cytosine base editor(CBE) and adenine base edit...The clustered regularly interspaced short palindromic repeats(CRISPR)–CRISPR-associated protein(Cas) system has been widely used for genome editing. In this system, the cytosine base editor(CBE) and adenine base editor(ABE) allow generating precise and irreversible base mutations in a programmable manner and have been used in many different types of cells and organisms. However, their applications are limited by low editing efficiency at certain genomic target sites or at specific target cytosine(C) or adenine(A) residues. Using a strategy of combining optimized synergistic core components, we developed a new multiplex super-assembled ABE(sABE) in rice that showed higher base-editing efficiency than previously developed ABEs. We also designed a new type of nuclear localization signal(NLS) comprising a FLAG epitope tag with four copies of a codon-optimized NLS(F4NLS^(r2)) to generate another ABE named F4NLS-sABE. This new NLS increased editing efficiency or edited additional A at several target sites. A new multiplex super-assembled CBE(sCBE) and F4NLS^(r2) involved F4NLS-sCBE were also created using the same strategy. F4NLS-sCBE was proven to be much more efficient than sCBE in rice. These optimized base editors will serve as powerful genome-editing tools for basic research or molecular breeding in rice and will provide a reference for the development of superior editing tools for other plants or animals.展开更多
Based on the oil,gas and water distribution characteristics of Khasib reservoir in Halfaya oilfield,Iraq,a core displacement experiment is designed to evaluate the influence of different displacement methods and displ...Based on the oil,gas and water distribution characteristics of Khasib reservoir in Halfaya oilfield,Iraq,a core displacement experiment is designed to evaluate the influence of different displacement methods and displacement parameters on oil displacement efficiency.The research shows that,in the displacement method with water injected from the edge of the reservoir,early depletion production is conducive to the elastic expansion of the gas cap,forming the three-dimensional displacement of"upper pressure and lower pushing",and the oil displacement effect is good.When gas injection at the top and water injection at the edge are used for synergistic displacement,the injection timing has different influences on the oil displacement effects of high and low parts.Considering the overall oil displacement efficiency,the injection pressure should be greater than the bubble point pressure of crude oil.Two displacement methods are recommended with the reasonable injection time at 20–25 MPa.The injection speed has the same influence on different injection media.Appropriately reducing the injection speed is conducive to the stability of the displacement front,delaying the breakthrough of injection media and improving the oil displacement effect.The reasonable injection rate of water flooding is 0.075 mL/min,the reasonable injection rates of water and gas are 0.15 mL/min and 0.10 mL/min,respectively in gas-water synergistic displacement.Gas-water synergistic displacement is conducive to the production of crude oil at high position,and has crude oil recovery 5.0%–14.8%higher than water flooding from the edge,so it is recommended as the development mode of Khasib reservoir at the middle and late stages.展开更多
基金supported by the Beijing Scholars Program[BSP041]。
文摘The clustered regularly interspaced short palindromic repeats(CRISPR)–CRISPR-associated protein(Cas) system has been widely used for genome editing. In this system, the cytosine base editor(CBE) and adenine base editor(ABE) allow generating precise and irreversible base mutations in a programmable manner and have been used in many different types of cells and organisms. However, their applications are limited by low editing efficiency at certain genomic target sites or at specific target cytosine(C) or adenine(A) residues. Using a strategy of combining optimized synergistic core components, we developed a new multiplex super-assembled ABE(sABE) in rice that showed higher base-editing efficiency than previously developed ABEs. We also designed a new type of nuclear localization signal(NLS) comprising a FLAG epitope tag with four copies of a codon-optimized NLS(F4NLS^(r2)) to generate another ABE named F4NLS-sABE. This new NLS increased editing efficiency or edited additional A at several target sites. A new multiplex super-assembled CBE(sCBE) and F4NLS^(r2) involved F4NLS-sCBE were also created using the same strategy. F4NLS-sCBE was proven to be much more efficient than sCBE in rice. These optimized base editors will serve as powerful genome-editing tools for basic research or molecular breeding in rice and will provide a reference for the development of superior editing tools for other plants or animals.
基金Supported by the Scientific Research and Technology Development Project of CNPC(2019D-4410)。
文摘Based on the oil,gas and water distribution characteristics of Khasib reservoir in Halfaya oilfield,Iraq,a core displacement experiment is designed to evaluate the influence of different displacement methods and displacement parameters on oil displacement efficiency.The research shows that,in the displacement method with water injected from the edge of the reservoir,early depletion production is conducive to the elastic expansion of the gas cap,forming the three-dimensional displacement of"upper pressure and lower pushing",and the oil displacement effect is good.When gas injection at the top and water injection at the edge are used for synergistic displacement,the injection timing has different influences on the oil displacement effects of high and low parts.Considering the overall oil displacement efficiency,the injection pressure should be greater than the bubble point pressure of crude oil.Two displacement methods are recommended with the reasonable injection time at 20–25 MPa.The injection speed has the same influence on different injection media.Appropriately reducing the injection speed is conducive to the stability of the displacement front,delaying the breakthrough of injection media and improving the oil displacement effect.The reasonable injection rate of water flooding is 0.075 mL/min,the reasonable injection rates of water and gas are 0.15 mL/min and 0.10 mL/min,respectively in gas-water synergistic displacement.Gas-water synergistic displacement is conducive to the production of crude oil at high position,and has crude oil recovery 5.0%–14.8%higher than water flooding from the edge,so it is recommended as the development mode of Khasib reservoir at the middle and late stages.
