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Genome editing

life science Maturity 11-13 Next article →

{

"text": "Genome editing, also known as genome engineering or gene editing, is a precise form of genetic engineering. This technology allows scientists to insert, delete, modify, or replace specific segments of DNA within the genome of a living organism. Unlike older genetic engineering methods, which often inserted new genetic material into a host genome at random locations, genome editing is site-specific. This means scientists can target exact locations to make changes. This precision helps reduce off-target effects, which are unintended changes to other parts of the DNA.

Possibilities of genome editing
Possibilities of genome editing

\n\nThe fundamental mechanism of genome editing relies on creating a double-strand break (DSB) in the target DNA. To achieve this, scientists use programmable nucleases, which act like molecular scissors. A nuclease consists of a DNA-binding domain (DBD) that recognizes a specific genomic locus and an effector domain that cuts the DNA. Once the nuclease creates a DSB, the cell naturally attempts to repair the damage through one of two major pathways. The first is non-homologous end joining (NHEJ), where enzymes directly join the broken DNA ends back together. The second is homology-directed repair (HDR), which is a more accurate process. In HDR, the cell uses a homologous sequence as a template to regenerate the missing DNA at the break point.

DsDNA break repair and CRISPR-based genome editing
DsDNA break repair and CRISPR-based genome editing

\n\nScientists have developed several distinct classes of engineered nucleases to perform these cuts. Meganucleases, discovered in the late 1980s, are naturally occurring enzymes that recognize very long DNA sequences, often between 14 and 40 base pairs. While they are highly specific, it is difficult to find a meganuclease that matches a specific desired sequence. To solve this, researchers use mutagenesis or computer models to create custom variants. Another class is Zinc finger nucleases (ZFNs). These use zinc finger motifs, which are small protein structures stabilized by zinc ions, to recognize short sequences of about 3 base pairs. By combining 6 to 8 of these fingers, scientists can target a specific sequence of approximately 20 base pairs.

Engineered Nucleases
Engineered Nucleases

\n\nAnother major class of tools is transcription activator-like effector nucleases (TALENs). Like ZFNs, TALENs use a non-specific DNA-cutting domain linked to specific DNA-recognizing peptides. The CRISPR-Cas system is perhaps the most famous modern method. Developed as a major advancement in 2015, the CRISPR/Cas9 system significantly improved the efficiency and practicality of large-scale genome editing. While earlier methods like meganucleases or ZFNs can be costly and time-consuming to design, CRISPR has made the process much more accessible. Researchers are even looking for alternatives, such as OMEGA proteins, which are endonucleases found in transposons.

MEGANUCLEASE-ZFN-TALEN-CRISPR-text-to-path
MEGANUCLEASE-ZFN-TALEN-CRISPR-text-to-path

\n\nThe history of this field shows a rapid progression in capability. Genetic engineering has been used to introduce new genetic elements since the 1970s. However, true genome editing began to emerge in the 1990s, though early efforts suffered from low efficiency. In 2011, Nature Methods named engineered nucleases as the Method of the Year. In 2015, Science magazine named CRISPR the Breakthrough of the Year. The field also faces significant ethical milestones. In 2019, the first humans were born from genome-edited embryos following a controversial affair involving scientist He Jiankui. This event led to calls for strict regulations regarding the manipulation of the human genome.

Overview of TALENs
Overview of TALENs

\n\nGenome editing has massive real-world significance across many industries. In agriculture, the Roslin Institute engineered pigs that are resistant to a virus causing porcine reproductive and respiratory syndrome. This disease costs pig farmers in the US and Europe approximately $2.6 billion every year. In food science, a tomato called Sicilian Rouge High GABA was approved for sale in Japan in 2020. This tomato contains more of an amino acid that may promote relaxation. In medicine, a 2020 US trial showed that CRISPR could be safely used on three cancer patients. These examples show how precise DNA changes can solve economic and health challenges.

Endogenous genes targeted
Endogenous genes targeted

\n\nBeyond simple edits, these technologies connect to complex biological systems and research. Scientists use homologous recombination (HR) to create transgenic mice with specific genes \"knocked out\" or "knocked in." This work was so impactful that Mario Capecchi, Martin Evans, and Oliver Smithies won the 2007 Nobel Prize in Physiology or Medicine. Researchers also use conditional targeting to study vital genes. By using systems like Cre-LoxP or Flp-FRT, they can switch genes on or off only in specific cells or at certain times. This allows for the study of gene functions without killing the organism prematurely. This deep level of control is essential for modern biological research and gene therapy.",

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🖼️ Images & Media (7)
File:MEGANUCLEASE-ZFN-TALEN-CRISPR-text-to-path.svg
MEGANUCLEASE-ZFN-TALEN-CRISPR-text-to-path.svg
File:DsDNA break repair and CRISPR-based genome editing.svg
DsDNA break repair and CRISPR-based...
File:Engineered Nucleases.jpg
Engineered Nucleases.jpg
File:Overview of TALENs.png
Overview of TALENs.png
File:Multiplex Automated Genomic Engineering (MAGE).png
Multiplex Automated Genomic Engineering (MAGE).png
File:Endogenous genes targeted.jpg
Endogenous genes targeted.jpg
File:Possibilities of genome editing.jpg
Possibilities of genome editing.jpg
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