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    • 5. 发明申请
    • METHOD FOR INTRODUCING NUCLEIC ACIDS INTO A CELL
    • 将核酸引入细胞的方法
    • WO2017191096A1
    • 2017-11-09
    • PCT/EP2017/060345
    • 2017-05-02
    • MILTENYI BIOTEC GMBH
    • JURK, MarionWILD, StefanBOSIO, Andreas
    • C12N15/87
    • C12N15/87C07K14/4703
    • The present invention provides the use of a nucleic acid encoding SOCS1 for enhancing the efficacy of introducing at least one nucleic acid of interest into a cell; a method of repeated transfection of a cell with at least one nucleic acid of interest comprising the steps of adding a) nucleic acid encoding SOCS1, and simultaneously or subsequently b) at least one nucleic acid of interest encoding at least one polypeptide of interest, wherein at least step b) is repeated at least once; and a method of electroporation of a cell with at least one nucleic acid of interest comprising the steps of adding to the cell a) a nucleic acid encoding SOCS1, and simultaneously or subsequently b) said at least one nucleic of interest. The at least one nucleic acid of interest and the nucleic acid encoding SOCS1 may be mRNAs, wherein each of said mRNAs has a poly(A) tail at its 3' end comprising at least 200 adenines.
    • 本发明提供编码SOCS1的核酸用于增强将至少一种感兴趣的核酸导入细胞的功效的用途; 用至少一种目标核酸重复转染细胞的方法,包括以下步骤:添加a)编码SOCS1的核酸,并且同时或随后b)编码至少一种目的多肽的至少一种感兴趣的核酸,其中 至少步骤b)重复至少一次; 以及用至少一种感兴趣的核酸对细胞进行电穿孔的方法,所述方法包括以下步骤:向细胞中加入a)编码SOCS1的核酸,并且同时或随后b)所述至少一种感兴趣的核酸。 至少一种目标核酸和编码SOCS1的核酸可以是mRNA,其中每种所述mRNA在其3'末端具有包含至少200个腺嘌呤的聚(A)尾巴。
    • 10. 发明申请
    • METHOD FOR THE OPTOINJECTION OF EXOGENOUS MATERIAL INTO A BIOLOGICAL CELL
    • 将外源材料插入生物细胞的方法
    • WO2017115220A1
    • 2017-07-06
    • PCT/IB2016/057829
    • 2016-12-20
    • FONDAZIONE ISTITUTO ITALIANO DI TECNOLOGIA
    • DIFATO, FrancescoSOLOPERTO, AlessandroPALAZZOLO, Gemma
    • C12N15/87C12N13/00C12M1/42B23K26/00
    • C12N13/00B23K26/00C12M35/02C12N15/87
    • Method for the optoinjection of exogenous material in a recipient biological cell enclosed by a membrane, which comprises: placing a biological cell on a planar surface of a substrate, transmitting a sub-ns pulsed laser beam through a variable convergence/divergence collimator, the collimator comprising a lens with focal length tunable by means of a variable amplitude control electrical signal; focusing the laser beam in a focal spot positioned along an axial direction substantially perpendicular to the substrate; moving the focal spot towards the cell along the axial direction by continuously varying the electric control signal from a first amplitude value corresponding to a first axial position zi of the focal spot above the apical surface of the cell to a second amplitude value, the second amplitude value defining a second focal length corresponding to a second axial position Zf of the focal spot, wherein the second amplitude value of the control signal is selected such that the second axial position is positioned inside the cell, at a second axial distance from the planar surface of the substrate, such that the focal spot traverses the membrane of the cell during the descent of the focal spot towards the cell producing a pore in the membrane, which allows the fluid solution containing exogenous material to enter the cell.
    • 用于在由膜包封的受体生物细胞中光注射外源性物质的方法,该方法包括:将生物细胞放置在基底的平坦表面上,将亚ns脉冲激光束透射通过 可变会聚/发散准直器,所述准直器包括焦距可借助于可变幅度控制电信号调谐的透镜; 将激光束聚焦在沿基本上垂直于衬底的轴向定位的焦点上; 通过连续地将电控制信号从对应于细胞顶点表面上方的焦点的第一轴向位置zi的第一幅度值改变为第二幅度值,将焦点沿着轴向方向移动到细胞,第二幅度 其定义对应于所述焦斑的第二轴向位置Zf的第二焦距,其中所述控制信号的所述第二幅值被选择为使得所述第二轴向位置位于所述单元内部,距离所述平坦表面 使得焦斑在焦斑下降过程中穿过细胞膜朝向在细胞膜中产生孔的细胞,这允许含有外源性物质的流体溶液进入细胞。