Site-specific cell perforation technique

Abstract

A technique for controlling membrane denaturation reactions other than physical shear force was developed. For example, the present invention provides, a method for causing membrane disruption at a specific site by reacting a stimulus such as light with a compound that is activated by the stimulus, where the reaction occurs on a membrane such as a biomembrane. It also provides a membrane structure such as cells in which a specific site has been disrupted, which are obtained by the present method. Introduction of substances such as genes also became possible by using this membrane structure. Further provided is a membrane-destroying member for disrupting a membrane at a specific site. Thus, the present invention enabled, for example, easy membrane penetration using components constituting microelectrodes, micromanipulators, and microinjectors, which were conventionally hardly usable in penetrating cell membranes.

Description

TECHNICAL FIELD [0001] The present invention relates to a method for perforating a membrane by partially treating the membrane (cell membrane, etc.) with a membrane-denaturing agent, etc. It also relates to a membrane-disrupting material having a membrane-denaturing effect. BACKGROUND ART [0002] In gene therapy and artificial substance production systems using living organisms, means of introducing a nucleic acid, a protein, and such, into the interior of a cell are extremely important. On the other hand, techniques for extracting structures such as the nucleus of a cell, are also gaining wide attention. In other words, it can be said that injecting and extracting substances into / from cells, the basic unit constituting organisms, is a fundamental technique of bioengineering. [0003] Conventional substance introduction techniques can be roughly categorized as follows: [0004] a) Introduction techniques targeting non-specific cell groups [0005] b) Introduction techniques targeting a spe...

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Benefits of technology

[0091] The present invention could also be utilized for functional electrical stimulation, and such, and for enhancing the accumulation/precision of stress-measuring therapeutic electrodes. As a part of rehabilitation medicine, a method called functional electrical stimulation is being used, in which metal electrodes are inserted into nerve bundles and electrical stimulation is given to improve nerve/muscle functions. At present, the method has not gone beyond stopped at setting up electrodes in a few places within the nerve bundle, and is insufficient in the aspects of site specificity/precision of the nerve stimulation. By conjugating the membrane perforation technique and already existing electrode accumulation technique, one-to-one joining of a nerve and electrode will become possible, and thereby, stimulating only the nerve that needs functional improvement could be accomplished.

[0092] The present invention could also be applied as a technique for transmitting signals from nerves to various artificial organs. Organs embedded within the body are controlled, not by direct neural information of the body, but strictly by indirect control. One example is artificial urethral valve control. These valves that are made by shape-memory metals, open by heating and close at normal body temperature. This problem is, that the closing and opening is controlled by an external heating device switch and cannot be directly controlled by the will of the patient. If stabilization/enhancement of precision of neural information becomes possible by neural interfaces using membrane-penetrating electrodes, these valves could be controlled as if they were a part of the body of the user. It is true that artificial urethral valves are used only a few times throughout the day, and the handling, the opening/closing, is also easy, and therefore, not much inconveniences are caused by switch manipulations outside the body in daily life. However, to control artificial organs that carry out more complicated acts in place of visceral functions, it is indispensable to make the control signal source be from autonomous nerves.

[0093] The present invention could also be used for connecting/controlling prosthetic hand/legs equipped with sensory organs and joints that are controllable similarly to the human body. At present, there is a remarkable enhancement in performance of externally powered prosthetic hands/legs for functional assistance following dismembering. However, the controlling information source utilizes myoelectric power left in the wearer in most cases, and the power is overwhelmingly short compared to the information amount originally needed to control the limbs. Also, the transmitting of sensation through the prosthetic hand/legs to the wearer is limited to just physical contact information via the area connected to prosthetic limb. Therefore, even if the handling of the prosthetic hands/legs is learned by training, the reality is that

Problems solved by technology

A fundamental problem underlying these cell treatments is the lack of techniques that could control the disruption of the cell membrane.

However, they could not respond to the demands of cytoengineering, which is to partially, and temporarily disrupt the cell membrane without causing cell death.

Also there were limitations to the method utilizing ph

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