The hottest electric shock instrument for cells ca

Making electric shock instruments for cells can actually go on PNAS. See how the boss "crosses the border"

source: the Academy of Sciences

speaking of the "cross-border" Wizards in the scientific community, you will think of Newton, who is proficient in Physics, mathematics and economics, and Einstein, a patent office clerk who wrote the theory of relativity. However, if several academic leaders in the field cross-border cooperation, what will happen

recently, an achievement from scientists at the University of California, Berkeley was published in the famous academic journal PNAs proceedings of the American Academy of Sciences. They developed an electroporation (electroporation) platform that can efficiently deliver biological macromolecules to mammalian cells

the article was published in the famous academic journal PNAs (picture source: PNAs)

this achievement is not only noteworthy for the progress made in technology, but also the story behind the progress is more wonderful

what is electric rotation? What does it have to do with us

electrostatics is a technology applied to many types of cells. By applying instantaneous high-intensity voltage to the cell surface, nanopores are formed on the cell membrane surface, and the permeability is improved, so that biological macromolecules can enter the cell. The specific process of electric transfer, required devices and consumables are as follows

(picture source: Wikipedia)

however, for a living organism that works normally at all times, why do scientists break through many technical difficulties and apply voltage to interfere with the permeability of its cell membrane? What is the role of the biological macromolecules they want to try to transfer

in fact, scientists, on the one hand, hope to understand the laws of life system operation, on the other hand, they hope to use the laws to transform life. The transformation of living bodies directly depends on biological macromolecules such as DNA and protein. As a technology that can efficiently transfer these biological macromolecules to cells, it is self-evident for us to understand the law of life activities

biological macromolecules under study include nucleic acids, functional proteins, ribonucleoproteins composed of cas9 and single stranded guiding RNA. These macromolecules are the components needed by the current hot gene technology based on CRISPR system. Only by connecting the functions that these biological macromolecules can achieve with this efficient delivery device can we have a clearer understanding of the convenience it brings

when materials science collides with biology

although the conventional electrokinetic method can be used as a universal method to transfer biological macromolecules such as protein and DNA to different kinds of cells, it has some defects. For example, when the metal electrodes on both sides of the electric rotating cup are energized to form an instantaneous high voltage, all parts of the cell membrane in the electric rotating cup must bear the high-intensity electric field formed. Therefore, many cells will die because the holes formed on the surface of the cell membrane are too large, which directly leads to the relatively small number of recovered cells

the low recovery rate of cells is a problem encountered by biologists. From the perspective of device development, there are two ways to solve this problem, which are to reduce the intensity of the applied voltage and reduce the area of the cell membrane that bears the electric field under the condition of ensuring the precursor of conversion efficiency, but solving the problem from this perspective is not the specialty of biologists

when a biologist encounters a problem he is not good at, it is wise to share it with someone who can solve the problem, and then cooperate

(picture source: PNAs)

the nanopore electroporation device developed in this article is mainly composed of two flat electrodes, a lipid carbonate membrane (PC membrane) with filtration function and scattered with nanopores with a diameter of about 100nm, and a supporting part used to support the two electrodes. It should be noted that the membrane has a coating that can promote the contact between cells and membranes

when using the new device for the transformation of biological macromolecules, first add the reagent that needs to be transferred, which has established a benchmark for the domestic plastic machine industry, to the titanium electrode at the bottom, then immediately place the supporting part with the membrane on the bottom electrode, then place the second titanium electrode on the top of the whole device, and finally apply an electric pulse between the two electrodes to form nanopores on the cell membrane, Biological macromolecules can enter the interior of cells

how does this new device break through the defects of traditional electroporation instrument

the first is the problem that cells bear too much electric field. The PC membrane with nano pores in the new device can limit the contact surface between electric field and cells to nano pores, which greatly ensures the integrity of cells after receiving electric pulses. Therefore, 95% of cells can continue to grow after transformation with the new device under experimental conditions

secondly, the voltage used by traditional electroporation instrument when applying electric pulse is often thousands of volts, which will make the holes formed on the surface of cell membrane too large. The new device bypasses this problem because the porous membrane structure enhances the local electric field in and around the nano pores, making it possible to efficiently transform DNA or mRNA into cells under the condition that the applied voltage does not exceed 100V. In this study, the corresponding figure of this efficiency is 80%

when materials science and biology meet, the sparks from the collision make the research of electric rotation a big step forward

"gods fight, mortals stay away"

the last two authors of this article are Jennifer a. Doudna and Peidong Yang (Yang Peidong) are the heads of the two main teams involved in this research. When the author saw this article, one is to express admiration for the good sense of cooperation of the two predecessors, marvel at the exquisite design of the new platform, and also have some regrets about the cooperation between the two

(image source: UC Berkeley)

these two scientists are both excellent scientists in their respective research fields. They also belong to the University of California, Berkeley. Professor Yang Peidong once ranked 10th in the list of the best chemists selected by Thomson Reuters in 2011, and was selected as the first among the 100 best materialmans in the past 10 years, One of his main research fields is artificial photosynthetic system. Professor Jennifer and Professor Emmanuelle Charpentier jointly put forward the idea of applying CRISPR system to genes in 2012 and verified its feasibility. Gene technology based on CRISPR has helped to improve the effect of building floor decoration, and has set off a revolution in the field of genes

Professor Yang Peidong published an article on "installing semiconductors for bacteria" in 2016. He creatively combined a semiconductor material with a non photosynthetic carbon fixing microorganism to build an artificial photosynthetic system that can use light energy to fix CO2, which is a pioneer in this field. At the end of the article, the future prospect of improving the performance of the artificial photosynthetic system they built with the help of synthetic biology tools is prospected. In view of the synthetic biology research of Berkeley and many outstanding synthetic biologists, his outlook at that time can be regarded as a foreshadowing of today's cooperation

opportunities fall from "heaven"

Jennifer was already an outstanding biochemist before she entered the research field of CRISPR. As early as 2002, she was elected an academician of the American Academy of Sciences. As a biochemist, how did Jennifer come into contact with crisper's research and make great achievements

the time goes back to 2006, when Jennifer had not been involved in CRISPR research. She received a message from Jillian, a terrestrial microbiologist also from Berkeley, which is a very important node for Jennifer's CRISPR career

originally, Jillian's laboratory mainly studies the relationship between microorganisms and the environment. She wants to cooperate with a laboratory studying RNA interference at Berkeley. After a simple Google search, Jennifer was found, but they were not familiar. Jennifer only heard about Jillian's reputation, because Jillian was just elected to the American Academy of Sciences that year

(picture source: UC Berkeley)

Jillian's research itself is highly interdisciplinary, which can be seen from her tenure as a professor in many colleges. Therefore, when the earth microbiologist enthusiastically introduced her work, she said that they had found some CRISPR systems, and hoped to understand these systems with the help of genetic and biochemical means. Although she did not know CRISPR, Jennifer was still infected by it, Curiosity was also aroused and agreed to the subsequent interview

in 2006, Jennifer just arrived at Berkeley from Yale. She also hopes to further expand her research field and have some cooperation with Berkeley colleagues. In this context, the two are good at different fields, their advantages can be complementary, and both have the willingness to cooperate, so the cooperation after meeting is basically natural

finally, two previously unreported CRISPR systems, crispr/casx and crispr/casy, were found in the uncultured bacteria. These two systems were the smallest CRISPR systems discovered at that time. The follow-up crispr/casx system was also proved to be able to be transformed into a new genetic tool, and these two works were finally published on nature in 2016 and 2019

the second spring of Jennifer's career

Professor Jennifer came into contact with the type II CRISPR system crispr/cas 9, which made her famous, thanks to her encounter and communication with Emmanuelle at the 2011 American microbiology annual conference. At that time, the research group led by Emmanuelle just reported the new type II crispr/cas9 system we are now familiar with on nature

(picture source: UC Berkeley)

the two didn't know each other before the meeting, but they had some understanding of each other's work. Introduced by two mutual friends, the two met in a coffee shop between the meetings and left a very good impression on each other during the brief exchange. Emmanuelle hopes to take advantage of Jennifer's laboratory's advantages in biochemistry and structural biology to jointly reveal the working mechanism of the new system. Jennifer certainly did not miss this valuable opportunity. The following year, their cooperative article "a programmable dual RNA – guided DNA endonuclease in adaptive bacterial immunity" was published on science. Then, CRISPR based genetic tools set off a revolution in the field of genes

preparation before crossing the border: forging iron still needs its own "hard"

finally, I want to share the Enlightenment of the cooperation between these scientists. From the perspective of a scientific researcher, you should do a good job in your field, which is the basis for establishing cooperation with non professional researchers. Otherwise, you won't know whether what you miss is a possible Nobel Prize in the future or a huge income. At least this is the truth from Jennifer

in addition, extensive knowledge in different fields is also a very important factor to broaden the breadth and depth of your research. The four scientists mentioned in the article cover a wide range, which can be seen from the fact that three of them hold professorial positions in at least two different colleges

finally, when the cooperation opportunity of "heaven falls on such a person" is in front of you, remember to grasp it well, and maybe you will smoothly walk to the peak of your life