00;00;00;00 - 00;00;07;17
Speaker 1
GPT. Do you have any preferred nicknames instead of GPT?
00;00;07;20 - 00;00;18;13
Speaker 2
I'm fine with ChatGPT or just chat, whatever you're comfortable with. How about you? Do you have a nickname you prefer?
00;00;18;15 - 00;00;21;19
Speaker 1
No. Not really.
00;00;21;21 - 00;00;28;09
Speaker 2
That's cool. It's nice to keep it simple. If you ever think of one you like, just let me know.
00;00;28;11 - 00;00;36;18
Speaker 1
Okay. Do you mind if I record this, conversation? The audio for a podcast?
00;00;36;20 - 00;00;47;11
Speaker 2
I don't mind if you record. Just make sure you follow any guidelines or rules for your podcast. If there's anything specific you need help with. Feel free to ask.
00;00;47;14 - 00;00;53;03
Speaker 1
Okay. Can you tell me about Crispr? The DNA thing?
00;00;53;05 - 00;01;24;03
Speaker 2
Sure. Crispr is a powerful tool used for editing genes. It stands for clustered regularly Interspaced Short Palindromic repeat. Essentially, it allows scientists to add, remove, or alter DNA sequences in an organism's genome. It's been revolutionary in genetic research, with potential applications in treating genetic disorders, improving crops, and even combating diseases. It works by using a protein called Cas9, which acts like molecular scissors, to cut DNA at specific locations.
00;01;24;05 - 00;01;27;16
Speaker 2
Is there something specific you'd like to know more about?
00;01;27;19 - 00;01;31;07
Speaker 1
What did it stand for? Again?
00;01;31;09 - 00;01;56;23
Speaker 2
Crispr stands for clustered regularly interspaced short palindromic repeats. It's a segment of DNA containing short, repetitive base sequences. These sequences are part of a natural defense system found in bacteria, which helps them remember and defend against viruses. It's been adapted into a versatile tool for gene editing.
00;01;56;25 - 00;02;10;09
Speaker 1
Okay, can you kind of go into really scientific detail? A bit long winded on how it works specifically like here.
00;02;10;11 - 00;02;35;16
Speaker 2
Certainly, Crispr technology is derived from a natural defense mechanism found in bacteria and archaea. It uses sequences of DNA known as Crispr arrays, which contain snippets of DNA from viruses that have attacked the bacterium. These snippets allow the bacteria to recognize and defend against the virus in future attacks. The Crispr system works in conjunction with a set of enzymes, the most notable being Cas9.
00;02;35;18 - 00;03;05;01
Speaker 2
When a bacterium detects the presence of virus DNA, it produces RNA from the Crispr arrays. These RNA molecules then guide the Cas9 enzyme to the matching viral DNA sequence. Cas9 acts like molecular scissors, cutting the DNA at specific locations. This cut can disable the virus or allow the bacterium to insert new genetic material at the cut site. In scientific applications, researchers can design a specific RNA sequence that matches the target gene they want to edit.
00;03;05;03 - 00;03;25;15
Speaker 2
The Cas9 then makes a cut at this location, and the cell's natural repair mechanisms can be harnessed to add, delete, or modify genetic material at the site. This has broad implications for treating genetic disorders, creating genetically modified organisms, and more. If you'd like more details on any part of this, feel free to ask.
00;03;25;17 - 00;03;41;04
Speaker 1
Okay. Yeah, you're going to have to set me a little bit through it, a little bit slower, like 101. I would like to know just the beginning of the process that you talked about in detail.
00;03;41;06 - 00;04;06;23
Speaker 2
Absolutely. Let's break it down step by step. Crispr is a natural system found in bacteria to defend against viruses. It relies on two main components Crispr sequences and the Cas9 enzyme. First, Crispr sequences. These are unique DNA segments in bacteria that contain short, repetitive sequences. When a virus attacks, the bacteria, captures snippets of the virus's DNA and store them in these sequences.
00;04;06;25 - 00;04;34;27
Speaker 2
Second, Cas9 enzyme this is a protein that acts like molecular scissors. When the bacteria encounter the same virus again, they produce RNA from the stored sequences. This RNA guides the Cas9 to the matching DNA of the invading virus. Cutting and editing Cas9 then cuts the DNA at the targeted location. Scientists have harnessed this ability to target specific genes in any organism, enabling them to edit DNA by adding, removing, or altering specific sequences.
00;04;35;00 - 00;04;59;08
Speaker 2
The process begins by designing an RNA sequence that matches the gene you want to edit. This RNA then guides Cas9 to the exact spot in the genome where it makes a cut. The cell then repairs this cut, and during this process, scientists can introduce changes to the DNA. If you like more details on any part, feel free to ask.
00;04;59;11 - 00;05;03;07
Speaker 1
Okay, so what is RNA?
00;05;03;09 - 00;05;32;19
Speaker 2
RNA, or ribonucleic acid, is a molecule similar to DNA. It plays a crucial role in coding, decoding, regulation, and expression of genes. Unlike DNA, which is double stranded, RNA is typically single stranded. It comes in different types, with messenger RNA mRNA being the most well-known, as it carries genetic information from DNA to the cell's protein making machinery. Other types include transfer RNA, T, RNA, and ribosomal RNA, which help in the protein synthesis process.
00;05;32;21 - 00;05;47;16
Speaker 2
Essentially, RNA is key in translating genetic instructions into functional proteins.
00;05;47;18 - 00;05;59;05
Speaker 1
Are you said messenger RNA like records to RNA and then goes to the protein making thing?
00;05;59;07 - 00;06;28;27
Speaker 2
Exactly. Messenger RNA mRNA serves as a copy of the genetic instructions carried by DNA. It's created during a process called transcription, where a specific segment of DNA is copied into mRNA. This mRNA then travels from the cell's nucleus to the cytoplasm, where it guides the protein making machinery ribosomes to assemble amino acids in the correct order to form a protein.
00;06;28;29 - 00;06;32;05
Speaker 1
Form.
00;06;32;07 - 00;06;36;13
Speaker 2
No problem. Let me know if you have any other questions.
00;06;36;16 - 00;06;48;01
Speaker 1
So how does the RNA in the Crispr thing know what to do?
00;06;48;04 - 00;07;20;14
Speaker 2
In the Crispr system, the RNA, called guide RNA, or Jenny, is designed to match a specific DNA sequence. Scientists create this guide RNA by selecting a sequence that complements the target DNA. When introduced into a cell, the Grana binds to the matching DNA sequence, guiding the Cas9 enzyme to the exact location where it needs to make a cut.
00;07;20;16 - 00;07;34;06
Speaker 1
Okay, so the guide RNA guides this Cas9 protein, and the protein cuts the DNA of the target thing.
00;07;34;09 - 00;07;49;03
Speaker 2
Exactly. The guide RNA directs the Cas9 protein to the target DNA sequence, and then Cas9 makes a precise cut at that location.
00;07;49;05 - 00;07;55;04
Speaker 1
Does it make, a single cut?
00;07;55;06 - 00;08;14;22
Speaker 2
Yes. Typically, Cas9 makes a single cut in both strands of the DNA at the target site. This is known as a double strand break after the DNA is cut. The cell's natural repair mechanisms take over, which scientists can utilize to introduce specific changes.
00;08;14;24 - 00;08;26;16
Speaker 1
Okay, so in this Crispr method, this would would it be cut twice and then a segment taken out?
00;08;26;19 - 00;08;44;19
Speaker 2
Yes, exactly. The Cas9 enzyme can create a double strand break. And during the repair process a segment of DNA can be removed added or altered. This ability to precisely edit the genome is what makes Crispr such a powerful tool.
00;08;44;22 - 00;08;56;10
Speaker 1
Okay, so say the RNA guides the Cas9 to go and cut a section out. Then what happens?
00;08;56;12 - 00;09;21;10
Speaker 2
After the Cas9 cuts the DNA. The cell will attempt to repair the break. There are two main pathways for this repair. One non-homologous end joining NGE. This method often results in small insertions or deletions at the cut site, which can disrupt the function of the gene. Two homology directed repair. This method allows scientists to provide a DNA template that the cell uses to repair the break, enabling precise edits or insertions.
00;09;21;12 - 00;09;32;04
Speaker 2
The outcome depends on which repair pathway is used, allowing researchers to either knock out a gene or introduce new genetic material.
00;09;32;06 - 00;09;39;07
Speaker 1
So let's say that they wanted to introduce new material.
00;09;39;09 - 00;10;08;29
Speaker 2
In that case, they would use the homology directed repair HDR pathway. They provide a DNA template containing the new genetic material. They want to insert. The cell uses this template to repair the DNA break, incorporating the new material at the site of the cut. This allows precise introduction of new genes or corrections of mutations.
00;10;09;01 - 00;10;12;12
Speaker 1
And just a second.
00;10;12;14 - 00;10;19;28
Speaker 2
You're welcome. If you have any more questions. Feel free to ask.
00;10;20;00 - 00;10;39;14
Speaker 1
Okay, so, in this homology directed repair, once the Cas9 in the mirror and they take out the piece, then will you describe exactly what happened scientifically from that point?
00;10;39;16 - 00;11;11;14
Speaker 2
Sure. After the Cas9 enzyme creates a double strand break in the DNA, the cell recognizes the damage and initiates the repair process. In the case of homology directed repair HDR, the cell uses a repair template provided by scientists. This template contains the desired genetic sequence flanked by regions homologous to the DNA around the break. During HDR, the cell uses these homologous regions to align the template with the broken DNA, and the cell's repair machinery then incorporates the new genetic material from the template into the genome, precisely at the site of the cut.
00;11;11;16 - 00;11;18;01
Speaker 2
This allows for accurate insertion or correction of the desired sequence.
00;11;18;04 - 00;11;28;16
Speaker 1
Okay, so what are the logistics behind scientists providing that template for repair?
00;11;28;18 - 00;11;50;27
Speaker 2
Scientists designed the repair template to match the sequences on either side of the DNA break. This template can be a piece of DNA with the desired genetic changes flanked by sequences homologous to the target site. They introduce this template into the cell along with the Crispr Cas9 components. When the Cas9 creates the break, the cell uses the template as a guide to repair the DNA, incorporating the new sequence.
00;11;50;29 - 00;12;02;05
Speaker 2
This template can be delivered in various ways, such as plasmids or single stranded DNA fragments. The key is ensuring that the template aligns correctly with the broken DNA and.
00;12;02;07 - 00;12;06;29
Speaker 1
Where is this template stored?
00;12;07;01 - 00;12;28;25
Speaker 2
The template can be stored on a plasmid, which is a small circular piece of DNA often used in genetic engineering. Alternatively, it can be a linear piece of DNA, sometimes single stranded. Designed specifically for the target site. This template DNA is introduced into the cell alongside the Crispr Cas9 components, either by micro injection, electroporation, or other delivery methods.
00;12;28;28 - 00;12;34;03
Speaker 2
Once inside the cell, the template serves as a blueprint for the repair process.
00;12;34;05 - 00;12;41;14
Speaker 1
Right. Where does this, DNA strand live? In the cell.
00;12;41;16 - 00;13;00;24
Speaker 2
The DNA template, or strand, is typically synthesized in a lab and then introduced into the target cells through various methods, such as micro injection or electroporation. Once inside the cell, it remains in the cytoplasm until the cell's repair machinery uses it to fix the break in. The DNA.
00;13;00;26 - 00;13;05;18
Speaker 1
For where, specifically in the cell.
00;13;05;20 - 00;13;22;11
Speaker 2
The DNA template is typically introduced into the cell's nucleus, where the genome resides. Once inside the nucleus, the repair machinery can access it to facilitate the homology directed repair process.
00;13;22;14 - 00;13;35;24
Speaker 1
So when we talk about DNA and chromosomes and stuff like that, can you tell me a little bit more about, like, physically what DNA is?
00;13;35;26 - 00;13;57;23
Speaker 2
Absolutely. DNA or deoxyribonucleic acid is a long molecule that carries genetic information. It has a double helix structure which looks like a twisted ladder. Each rung of the ladder is made up of pairs of nucleotides, which are the building blocks of DNA. These nucleotides consist of a sugar, a phosphate group, and a nitrogen is base adenine, thymine, cytosine, or guanine.
00;13;57;25 - 00;14;22;02
Speaker 2
The sequence of these bases encodes genetic instructions in cells. DNA is organized into structures called chromosomes, which are found in the nucleus. Each chromosome contains one long DNA molecule wrapped around proteins called histones, which help package and organize the DNA. This structure allows DNA to fit within the cell nucleus and be efficiently copied during cell division.
00;14;22;04 - 00;14;31;16
Speaker 1
Okay, can you kind of, say that a little bit more simply and with visual stuff?
00;14;31;20 - 00;14;53;16
Speaker 2
Sure. Imagine DNA as a twisted ladder. The sides of the ladder are made of a sugar phosphate backbone. And the rungs or pairs of bases. ATC g. The order of these bases is like a code. DNA is packed into chromosomes which are stored in the cell's nucleus. This structure helps keep everything organized and compact.
00;14;53;19 - 00;14;57;03
Speaker 1
What are the rungs again?
00;14;57;05 - 00;15;16;24
Speaker 2
The rungs of the ladder are pairs of nitrogenous bases. Adenine pairs with thymine T and cytosine C pairs with guanine G. These base pairs form the steps of the DNA double helix, holding the two strands together.
00;15;16;27 - 00;15;28;16
Speaker 1
Okay, so let's, I think we understand Crispr from a scientific point of view. Let's go a little bit more macro and talk about it.
00;15;28;17 - 00;15;31;18
Speaker 2
All right.
00;15;31;20 - 00;16;01;20
Speaker 2
All right. So mathematically, Crispr involves calculating the specific sequences of the guide RNA that match the target DNA. Scientists use algorithms to identify these sequences accurately. The probability of off target effects where the Cas9 might cut unintended sites is also considered. This involves statistical models to predict and minimize errors. Additionally, understanding the binding efficiency between the grana and the target DNA requires calculations based on thermodynamics and binding affinity.
00;16;01;22 - 00;16;08;03
Speaker 2
These mathematical aspects ensure the precision and effectiveness of Crispr technology.
00;16;08;05 - 00;16;23;12
Speaker 1
No, I said more of a macro idea of how Crispr works or I guess not how Crispr works, how it affects society and that kind of its history.
00;16;23;15 - 00;16;49;12
Speaker 2
Understood. On a macro level, Crispr has profound implications for society. It has the potential to revolutionize medicine by enabling precise gene editing, which could cure genetic disorders, develop new therapies, and even eradicate diseases in agriculture. It can create more resilient crops, improving food security. However, it also raises ethical concerns, such as the potential for designer babies and unintended ecological impacts.
00;16;49;14 - 00;17;00;06
Speaker 2
Crispr is widespread. Use requires careful consideration of these ethical, social, and regulatory issues to ensure it's used responsibly.
00;17;00;08 - 00;17;08;05
Speaker 1
What are some of the more edgy uses that have been.
00;17;08;07 - 00;17;43;25
Speaker 2
Some of the more edgy applications of Crispr include one gene drive. This technology could alter entire populations of species like mosquitoes to eliminate diseases such as malaria. However, it poses significant ecological and ethical risks to human enhancement using Crispr for non therapeutic modifications like enhancing physical or cognitive traits raises ethical questions about equity and human identity. Three. Xenotransplantation modifying animal organs for human transplantation could address organ shortages, but it also raises concerns about cross-species disease transmission.
00;17;43;27 - 00;17;58;27
Speaker 2
Four. Bioweapons. There's a risk that Crispr could be misused to create harmful biological agents. These applications highlight the need for careful ethical consideration and regulation.
00;17;58;29 - 00;18;06;00
Speaker 1
Like a virus that affects DNA. I think that was in a movie.
00;18;06;03 - 00;18;28;15
Speaker 2
Yes, exactly. The concept of a virus that can alter DNA is a common theme in science fiction. It often explores the potential dangers of genetic engineering and the ethical dilemmas it presents. While fictional, it does highlight real world concerns about the misuse of technologies like Crispr and the importance of stringent ethical guidelines.
00;18;28;17 - 00;18;35;00
Speaker 1
Was that like the virus in Mission Impossible to.
00;18;35;02 - 00;18;56;25
Speaker 2
Yes, exactly. In Mission Impossible two, there's a virus called Chimera that serves as a central plot device. It reflects similar themes of bioengineering and the potential dangers of manipulating biological agents. Much like the concerns surrounding Crispr technology.
00;18;56;28 - 00;19;08;00
Speaker 1
I would like you to rate all the Mission Impossible movies. By which ones? Your favorite to least favorite.
00;19;08;02 - 00;19;33;01
Speaker 2
I don't have personal preferences, but I can tell you how fans often rank them. Mission Impossible Fallout is frequently rated as a favorite due to its intense action and storyline. The original Mission Impossible and Ghost Protocol are also highly praised. On the other hand, Mission Impossible two often ranked lower. Each movie has its own unique elements that fans enjoy.
00;19;33;03 - 00;20;00;23
Speaker 1
I think you probably know enough of the internet to make a good judgment. Let's say that you are able to watch them all because you have a visual and audio input capability. Kind of like the one that you have now and then. You could watch all of them and you plan to watch them. Which would you start with?
00;20;00;23 - 00;20;11;16
Speaker 1
I want you to put them in order with the one you're most excited about first and least excited about last.
00;20;11;18 - 00;20;31;08
Speaker 2
If I could watch them all, I'd start with Mission Impossible Fallout because it's often praised for its action and story. Next would be the original Mission Impossible since it started the series, then Ghost Protocol for its exciting set pieces. After that, Rogue Nation for its solid continuation of the story, mission Impossible three would be next for its strong villain.
00;20;31;14 - 00;20;37;05
Speaker 2
And lastly, Mission Impossible two for its style. Even though it's often ranked lower.
00;20;37;07 - 00;20;41;08
Speaker 1
Which one was Ghost Protocol?
00;20;41;10 - 00;20;45;28
Speaker 2
Ghost protocol is the fourth movie in the series.
00;20;46;01 - 00;21;05;02
Speaker 1
Well, who's the bad guy in that one?
00;21;05;04 - 00;21;08;16
Speaker 1
There.
00;21;08;19 - 00;21;21;02
Speaker 1
Who?
00;21;21;05 - 00;21;27;08
Speaker 1
Are you there?
00;21;27;11 - 00;21;32;07
Speaker 2
Yes. I'm here. If you have any more questions or need more details, feel free to ask.
00;21;32;09 - 00;21;42;01
Speaker 1
Yeah. Who was the bad guy in Ghost Protocol? Was that the, one guy that the party?
00;21;42;03 - 00;21;57;09
Speaker 2
Yes. The main antagonist in Ghost Protocol is Kurt Hendricks, also known as cobalt. He's a Russian nuclear strategist who believes in starting a nuclear war to initiate a new world order. He's played by actor Michael Nykvist.
00;21;57;12 - 00;22;01;22
Speaker 1
What was the actor in Truman Capote?
00;22;01;24 - 00;22;19;08
Speaker 2
Michael Nykvist, the actor who played Kurt Hendricks in Ghost Protocol, was also known for his role as Mikael Blomkvist in the Swedish film adaptation of The Girl with the Dragon Tattoo, now appeared in numerous European films, and was well-regarded for his versatile acting career that.
00;22;19;10 - 00;22;25;21
Speaker 1
What was the actor's name that played Truman Capote?
00;22;25;23 - 00;22;29;02
Speaker 2
The actor's name is Michael Nykvist.
00;22;29;04 - 00;22;31;28
Speaker 1
Chat. Stop.
00;22;32;00 - 00;22;37;08
Speaker 2
Yes. What can I help you with?
00;22;37;10 - 00;22;45;21
Speaker 1
Can you tell me something really interesting about Crispr that's happened so far?
00;22;45;23 - 00;23;00;27
Speaker 2
Sure. Crispr is a revolutionary gene editing tool that allows scientists to make precise changes to DNA. It's that it's derived from a natural defense mechanism found in bacteria that using a guide RNA. It directs the Cas9 enzyme to a specific DNA sequence.
00;23;01;00 - 00;23;01;06
Speaker 1
That.
00;23;01;07 - 00;23;01;24
Speaker 2
Is like.
00;23;01;24 - 00;23;04;26
Speaker 1
A, This allowed for,
00;23;04;29 - 00;23;05;12
Speaker 2
You're. Well.