Basic Research on Effectiveness of Vibratory Microinjection Method

振动显微注射法有效性的基础研究

基本信息

  • 批准号:
    16500281
  • 负责人:
  • 金额:
    $ 1.98万
  • 依托单位:
  • 依托单位国家:
    日本
  • 项目类别:
    Grant-in-Aid for Scientific Research (C)
  • 财政年份:
    2004
  • 资助国家:
    日本
  • 起止时间:
    2004 至 2006
  • 项目状态:
    已结题

项目摘要

To increase the production rate of transgenic animals, we developed Vibratory Microinjection System (VMS). This system was aimed at facilitating gene transfer by vibrating a micropipette longitudinally during insertion into cells.1.The VMS providing any frequency less than 18 kHz was evaluated. The vibratory microinjection (VM) at several frequencies was compared with the ordinary (non-vibratory) microinjection (OM). Fertilized eggs obtained from BDF-1 mice were equally allotted to VM and OM groups. The eggs in each group were divided into some sets of 30 eggs. The 30 eggs in each set were injected with Venus genes using a single micropipette.(1)Compared with OM, VM enabled micropipettes to penetrate zona pellucida with significantly less cellular deformation rates, reducing them by about 20%. (2)After four-day culture of eggs having survived the injection, the VM group showed 2 to 3 times higher rates of development to the blastocyst stage than the OM group did, although it did slightly higher death rates (NS). (3)‘Pulling-out' event, i.e., an event that a micropipette pulled out a part of nuclear DNA before finishing the given number of injections, took place significantly less frequently in VM group than in the OM group. The pulling-out event usually necessitates replacement of the micropipette with a new one.2.We developed ultrasonic VMS (UVMS) with upper limit of frequency of 100kHz. We conducted safety evaluation of UVMS, and confirmed no cellular damage caused by ultrasonic vibration : extreme change in intracellular pressure, cavitation or fragmentation of DNA.In conclusion, VMS achieved zygote injection with significantly less cellular deformation and also resulted in significantly better embryonic development, suggesting improvement in production of transgenic animals. VMS can also cut the cost of micropipettes and save the trouble of frequent replacement of micropipettes.
为了提高转基因动物的生产率,我们开发了振动显微注射系统(VMS)。该系统旨在通过在插入细胞期间纵向振动微量移液器来促进基因转移。1.评估提供小于18kHz的任何频率的VMS。将多个频率下的振动显微注射(VM)与普通(非振动)显微注射(OM)进行比较。从 BDF-1 小鼠获得的受精卵平均分配给 VM 组和 OM 组。每组的鸡蛋被分成若干组,每组 30 个鸡蛋。每组30个卵子使用单个微量移液器注射维纳斯基因。(1)与OM相比,VM使微量移液器穿透透明带的细胞变形率显着降低,减少了约20%。 (2)注射后存活的卵子培养四天后,VM组的囊胚期发育率比OM组高2至3倍,尽管其死亡率(NS)略高。 (3)‘拉出’事件,即在完成给定次数的注射之前微量移液器拉出部分核DNA的事件,在VM组中发生的频率明显低于OM组。拔出事件通常需要更换新的微量移液器。2.我们开发了超声波VMS(UVMS),频率上限为100kHz。我们对UVMS进行了安全性评估,并确认超声波振动不会造成细胞损伤:细胞内压力的极端变化、空化或DNA断裂。总之,VMS实现了受精卵注射,细胞变形明显减少,并且胚胎发育明显更好,这表明转基因动物的生产得到改善。 VMS还可以降低微量移液器的成本,省去频繁更换微量移液器的麻烦。

项目成果

期刊论文数量(28)
专著数量(0)
科研奖励数量(0)
会议论文数量(0)
专利数量(0)
Development of Vibratory Microinjection System to increas production rate of transgenic animals
开发振动显微注射系统以提高转基因动物的生产率
  • DOI:
  • 发表时间:
    2006
  • 期刊:
  • 影响因子:
    0
  • 作者:
    Miyawaki F;Hasegawa J;Kikuchi T;Tsuchiya M;Saito Y;Suzuki H;Ippongi T;Kobayashi K;Itohara S;Miyawaki F et al.
  • 通讯作者:
    Miyawaki F et al.
振動型マイクロインジェクション装置
振动显微注射装置
  • DOI:
  • 发表时间:
    2005
  • 期刊:
  • 影响因子:
    0
  • 作者:
  • 通讯作者:
Development of Vibratory Microinjection System to increase production rate of transgenic animals
开发振动显微注射系统以提高转基因动物的生产率
Effects of Vibratory Microinjection on Mouse Zygotes and Subsequent In vitro Development.
振动显微注射对小鼠受精卵及其后续体外发育的影响。
  • DOI:
  • 发表时间:
    2005
  • 期刊:
  • 影响因子:
    0
  • 作者:
    Miyawaki F;Hasegawa J;Kikuchi T;Tsuchiya M;Saito Y;Suzuki H;Ippongi T;Kobayashi K;Itohara S;Miyawaki F et al.;菊池智行 他;Miyawaki F et al.
  • 通讯作者:
    Miyawaki F et al.
Vibration Type Micro Injection Device
振动式微注射装置
  • DOI:
  • 发表时间:
    2006
  • 期刊:
  • 影响因子:
    0
  • 作者:
  • 通讯作者:
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MIYAWAKI Fujio其他文献

MIYAWAKI Fujio的其他文献

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{{ truncateString('MIYAWAKI Fujio', 18)}}的其他基金

Development and systematization of Scrub Nurse Robot toward clinical use
临床应用的擦洗护士机器人的开发和系统化
  • 批准号:
    19300186
  • 财政年份:
    2007
  • 资助金额:
    $ 1.98万
  • 项目类别:
    Grant-in-Aid for Scientific Research (B)
Investigation of Usefulness of "Recovery-Directed Left Ventricular Assist Device" invented based on a new concept
基于新概念发明的“恢复导向左心室辅助装置”的实用性研究
  • 批准号:
    13480299
  • 财政年份:
    2001
  • 资助金额:
    $ 1.98万
  • 项目类别:
    Grant-in-Aid for Scientific Research (B)
Development of New Circulatory Assist Device-Split-Circulation Assits Device
新型循环辅助装置-分流循环辅助装置的研制
  • 批准号:
    08558104
  • 财政年份:
    1996
  • 资助金额:
    $ 1.98万
  • 项目类别:
    Grant-in-Aid for Scientific Research (A)
Basic study for the development of small caliber vascular prosthesis with alternately aligned static magnetic field
交替排列静磁场小口径人工血管研制的基础研究
  • 批准号:
    03807089
  • 财政年份:
    1991
  • 资助金额:
    $ 1.98万
  • 项目类别:
    Grant-in-Aid for General Scientific Research (C)

相似海外基金

Earthquake Induced Longitudinal Vibration of Earth Dams
地震引起的土坝纵向振动
  • 批准号:
    8005615
  • 财政年份:
    1980
  • 资助金额:
    $ 1.98万
  • 项目类别:
    Standard Grant
Earthquake Induced Longitudinal Vibration of Earth Dams
地震引起的土坝纵向振动
  • 批准号:
    7822865
  • 财政年份:
    1979
  • 资助金额:
    $ 1.98万
  • 项目类别:
    Standard Grant
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