top student at medical school

Chapter 212 Gene Editing Technology (First Update, Please Subscribe)
Animal Basic Experimental Technology Package Level 5 (0/500000).

The color is slightly different from the previous fluorescent green; it has become a pale gold.

Although there were no particularly dazzling or eye-catching changes, Fang Ziye, as the skill holder, knew that he had now mastered a great deal of content related to basic animal experiments.

Animal basic experimental techniques are not necessarily basic, but rather a basic methodology, and the basis for the subjects being operated on, rather than the operational techniques being very basic and something anyone can do.

Skills are the mastery of methodology, but mastering methodology does not mean that one has completely mastered all animal experiments. One still needs to rely on the most basic principles and ideas to complete the design of animal experiments.

Establishing animal models is just one of many animal experiments.

Fang Ziye knew long ago that the methodology used to establish animal models is called gene editing.

The explanation of gene editing technology is very simple!
This refers to the process of modifying specific targets in an organism's genome. It efficiently and precisely achieves gene insertion, deletion, or replacement, thereby altering its genetic information and phenotypic characteristics.

However, putting it into practice is quite difficult.

After all, gene editing technology can be broadly divided into gene knockout and gene knock-in, which is similar to silencing and overexpressing a gene in cell experiments.

Its basic principle is very simple...

Fang Ziye didn't think much about it, but rather wondered how this operational technique could bring him significant benefits in completing his current experimental work.

The answer is obvious!

The design of the experimental direction must precede the completion of the experimental operation.

One is theoretical understanding, and the other is practical application.

Pre-designing is like knowing a place exists, while completing an experiment means you actually go to the place you want to go. These are two different concepts.

The design of the experiment does not actually require you to have super high operational skills, but rather your ability to collect and summarize data, and then to develop a general process.

Whether or not it can be completed depends on individual ability.

Anyone with even a little scientific research experience knows that everyone can imagine a technological direction that will shock the world, but whether that imagination can be realized is another matter.

For example, building a ladder to the moon, tiling the Great Wall, and constructing a swimming pool in the Pacific Ocean.

Fang Ziye had designed experimental directions a long time ago, one of which was the establishment of the HK2 animal gene knockout model.

Rereading the notebook and slowly rereading the experimental design from back then made me feel much calmer.

Fang Ziye no longer felt particularly excited or thrilled. On the contrary, the improvement in his technical skills had calmed his mind and made him feel much more at peace.

Fang Ziye didn't know why either!
Just a few minutes ago, I really wanted to finish this project and see what that senior colleague was really like, but after my experimental skills improved, I didn't feel that way anymore.

Fang Ziye guessed that perhaps, compared to rendering that senior uncle powerless in front of his master, the improvement of his own abilities and the difference in strength gave Fang Ziye a greater sense of accomplishment.

Rather than seeing the true colors of certain people, Fang Ziye preferred to truly experience the joy of experimental results.

The ink on the notes written with a ballpoint pen has dried.

Experimental design and planning:
Define the scientific objectives and questions of the research. Develop an experimental plan, including the selection of animal models and gene knockout methods.

Selection of animal models:

The selection of appropriate experimental animals, typically mice or rats, depends on the research question and the feasibility of the model.

These two points are completely copied from the basic theory section and are just the general outline of the project experiment, so they don't have any special significance.

More importantly, it's below.

Determine the specific method for HK2 gene knockout!
Common methods include CRISPR/Cas9 technology and embryonic stem cells from gene knockout mice.

With just one sentence, the difficulty level suddenly increased.

CRISPR/Cas9 technology is a gene-editing technology. Editing genes requires this technology, but how to knock out genes and obtain embryonic stem cells from gene-injected mice after the knockout process becomes a mystery.

The fact that experimental animals cannot reproduce after animal experiments does not mean that the experimental animal model was a complete organism from the beginning.

This requires removing the embryonic stem cells from mice after fertilization and performing gene editing.

The following steps are:

Designing gene editing tools and conducting in vitro gene editing experiments:
Gene editing was performed in vitro on embryonic stem cells or other suitable cells to verify the effectiveness of the editing tool.

In vivo gene editing experiments:

Edited embryonic stem cells or gene-editing tools are introduced into animals, typically mouse or rat embryos.

Screening for normal gene-edited variants:
Animals with the HK2 gene successfully knocked out were screened using molecular biology techniques such as PCR and Southern blotting.

Animal reproduction:

Animals with completely knocked-out genes are created using successfully edited embryos, typically through breeding and selection to produce gene knockout offspring. Verification of gene knockout effectiveness:

Detailed molecular biological and biochemical analyses were performed on the generated gene knockouts to confirm whether the HK2 gene was successfully knocked out.

These experiments, which seemed impossible and extremely difficult, now seem less challenging to Fang Ziye when he reviews them one by one.

At least now, they don't seem like the empty shells or dreamlike illusions I used to see; they seem a little more substantial.

This is because Fang Ziye obtained feedback on the feasibility of the experimental operation through his own abilities.

If you've decided to do it, then go for it.

Each step needs to be detailed. For example, you can contact a large animal testing company and ask if they can provide embryonic stem cell extraction technology. If they cannot, can they provide the platform and address for the experimental operation so that you can operate it yourself?

If it can be provided, then how they transport it and where it is transported are all questions.

This is no ordinary animal experiment that can be done in any animal laboratory. Fang Ziye suspects that he may have to return to Wuhan, or even fly to Shanghai, Beijing, or Guangzhou, places with developed economies and advanced medical technology that are at the forefront of scientific research in China.

But no matter what, everything is difficult at the beginning. Having an idea at least gives you the confidence to do it.

Fang Ziye contacted the large animal testing center in Wuhan that his master, Yuan Weihong, had taken him to before.

The staff member immediately said they would ask.

A few minutes later, the person named Xiaoya contacted her and said, "Professor Fang, isn't your question a bit too advanced? Our technical director, who is also our company's PhD, said that your experimental direction is not something our lab can handle."

"Teacher Fang, we have other things here..."

Fang Ziye read half of it and then politely declined her further suggestions.

If it were completely easy, why would I consult you? Wouldn't it be better for me to just come and do it myself?

After consulting with representatives from several other experimental centers, it was discovered that there was only one animal laboratory in Wuhan capable of performing such operations, and that laboratory was located within the internal laboratory of Tongji Hospital affiliated with Huazhong University of Science and Technology.

"Professor Fang, the animal laboratory team at Tongji Hospital, in principle, only accepts advanced animal experiments from its own staff and does not conduct such experiments outside the hospital."

"Of course, if you need our help, Professor Fang, we can try to make some contact," replied Xiaohui, the manager of a biotechnology company, with great enthusiasm.

If we can achieve results in Fang Ziye's area, it will at least surpass the team's performance, possibly even in a year.

If you want to create new animal models, you could need tens of millions, or less than two million, but you don't even need to try.

"Yes, I just wanted to ask a question first. Thank you, Professor Wang. We can contact each other if needed," Fang Ziye said.

All he needs to do is get the address. Whether he needs to get in through a teacher in the college or through other channels is another matter.

If there's absolutely no other way, then you should ask the other party for help. That's the right thing to do. Otherwise, they'll hold you hostage and you'll end up owing them a favor. If you don't buy their things, that's really inappropriate.
But what if the other company's products aren't suitable?

Tongji Hospital!

If this were before the time of Xia En City, Fang Ziye could only ask his teacher for help and see if he had any connections. But now, Fang Ziye can ask his senior brother Wu Xuanqi.

In addition, Fang Ziye also knew that there were at least three experimental centers in Shanghai that could perform such operations, but at present, there were very few teams that would think of performing such high-end and cutting-edge animal experiments with extremely high uncertainty.

……

The next day.

After checking the rooms, Fang Ziye chatted casually with Wu Xuanqi for a few minutes.

Wu Xuanqi's first reply was: "..."

The reply was: "Ziye, can we not be so discouraging? You're asking about gene editing technology, you're getting a bit too advanced."

"Are there any good research topics?"

"Well, senior brother, we're also planning to try establishing animal models for animal experiments, to make some money, and to pave the way," Fang Ziye replied, not entirely sure.

Within the province and across the country, there are at least 800 to 1,000 teams that want to conduct ultra-high-end experiments.

Who hasn't had a dream?

But how many teams actually succeed in the end? The vast majority of teams' operations end in failure, essentially throwing money down the drain. Yet, there are still countless teams that will continue to waste their money.

"Ziye, let's be honest. Are you serious about this, or do you just want to have some fun?"

“If you’re sincere, we’ll follow the sincere procedure; if you just want to have some fun, we’ll follow the fun procedure,” Wu Xuanqi said enthusiastically.

"Senior brother, are there any differences between them?" Fang Ziye pressed.

(End of this chapter)