Synthetic guide RNA (sgRNA) is a type of RNA molecule that is artificially designed and created for use in genome editing techniques, particularly in the context of CRISPR-Cas9 gene editing.
Here's how synthetic guide RNA works:
1. Target Identification: Scientists identify the specific gene or genomic region they want to modify.
2. Design and Synthesis: Based on the target DNA sequence, they design a synthetic guide RNA that is complementary to the DNA sequence near the target site. This guide RNA includes a region that matches the target DNA and a region that interacts with the Cas9 protein.
3. Delivery: The synthetic guide RNA is introduced into the target cells, often along with the Cas9 protein or the Cas9 gene.
4. Guiding Cas9:Once inside the cell, the synthetic guide RNA binds to the Cas9 protein, forming a complex.
5. DNA Cleavage: The Cas9 protein, guided by the synthetic RNA, searches for the complementary DNA sequence in the genome. When it finds a match, it induces a double-strand break (DSB) at that precise location.
6. Repair Mechanisms: Cellular repair mechanisms are then activated to fix the DSB. This can result in various outcomes, including gene knockout (if the repair process introduces errors) or gene correction (if a DNA template is provided along with the synthetic guide RNA and Cas9 to facilitate precise repair).
Synthetic guide RNAs are customized to target specific genes or regions of interest, making them a powerful tool in genetic engineering and genome editing applications. They offer the ability to manipulate and study genes with a high degree of specificity and precision.
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This online tool shows commonly used genetic codon frequency table in expression host organisms including Escherichia coli and other common host organisms.
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