Difference between revisions of "Collections/CRISPR-Cas for Diagnosis"
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<h3>1. CRISPR-Cas12/13 as detection systems</h3> | <h3>1. CRISPR-Cas12/13 as detection systems</h3> | ||
<p>Recently (ends of 2021), especially due to the COVID-19 pandemic situation, <b>fast diagnostic procedures</b> based on CRISPR-Cas with the use of different Cas12 and Cas13 protein species, have gained a lot of relevance because of having proved to successfully work [1][2][3]. That is why ARIA thinks that the CRISPR-Cas for Diagnosis Part Collection might be widely useful for <b>future iGEM teams</b> making use of this emergent application of the technology.</p> | <p>Recently (ends of 2021), especially due to the COVID-19 pandemic situation, <b>fast diagnostic procedures</b> based on CRISPR-Cas with the use of different Cas12 and Cas13 protein species, have gained a lot of relevance because of having proved to successfully work [1][2][3]. That is why ARIA thinks that the CRISPR-Cas for Diagnosis Part Collection might be widely useful for <b>future iGEM teams</b> making use of this emergent application of the technology.</p> | ||
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<p>The principles behind the detection of a specific target DNA/RNA fragment by these two types of Cas proteins (12/13, respectively) are simple. They are based on the <b>emergence of a fluorescent signal</b> due to the <b>collateral trans cleavage activity of these Cas nucleases</b>, which means that when they get activated by the presence of the target fragment of genetic material, they not only cut it but also cut any other DNA which is present in the media [refs]. By taking advantage of this property, any desired target sample can be detected simply by introducing with it a <b>reporter</b> which releases a fluorophore when being cut, producing in this way a fluorescent signal that can be measured with a fluorescence detector.</p> | <p>The principles behind the detection of a specific target DNA/RNA fragment by these two types of Cas proteins (12/13, respectively) are simple. They are based on the <b>emergence of a fluorescent signal</b> due to the <b>collateral trans cleavage activity of these Cas nucleases</b>, which means that when they get activated by the presence of the target fragment of genetic material, they not only cut it but also cut any other DNA which is present in the media [refs]. By taking advantage of this property, any desired target sample can be detected simply by introducing with it a <b>reporter</b> which releases a fluorophore when being cut, producing in this way a fluorescent signal that can be measured with a fluorescence detector.</p> | ||
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<h3>2. Part Collection</h3> | <h3>2. Part Collection</h3> | ||
<p>This part collection is created with the purpose of helping other future iGEM teams for the guidance in the design of their parts needed to accomplish the full detection system described above. For that, we have created and documented it in a way that it is <b>easy-to-use</b> and <b>user-friendly</b>, as it is as much structured and organized as possible, being all the part pages linked between them in a way that an external user can comfortably replicate our design.</p> | <p>This part collection is created with the purpose of helping other future iGEM teams for the guidance in the design of their parts needed to accomplish the full detection system described above. For that, we have created and documented it in a way that it is <b>easy-to-use</b> and <b>user-friendly</b>, as it is as much structured and organized as possible, being all the part pages linked between them in a way that an external user can comfortably replicate our design.</p> | ||
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<h4 style="margin-left:15px;"> - Categories</h4> | <h4 style="margin-left:15px;"> - Categories</h4> | ||
Revision as of 22:59, 16 October 2021
This part collection aims to nest all these parts needed for the CRISPR-Cas technology to be used as a detection system with diagnostic purposes. The creators of this collection are ARIA, the 2020 UPF Barcelona iGEM team, who make use of the CRISPR-Cas technology for detecting antibiotic resistance genes in, as a proof-of-concept, laboratory samples.
1. CRISPR-Cas12/13 as detection systems
Recently (ends of 2021), especially due to the COVID-19 pandemic situation, fast diagnostic procedures based on CRISPR-Cas with the use of different Cas12 and Cas13 protein species, have gained a lot of relevance because of having proved to successfully work [1][2][3]. That is why ARIA thinks that the CRISPR-Cas for Diagnosis Part Collection might be widely useful for future iGEM teams making use of this emergent application of the technology.
The principles behind the detection of a specific target DNA/RNA fragment by these two types of Cas proteins (12/13, respectively) are simple. They are based on the emergence of a fluorescent signal due to the collateral trans cleavage activity of these Cas nucleases, which means that when they get activated by the presence of the target fragment of genetic material, they not only cut it but also cut any other DNA which is present in the media [refs]. By taking advantage of this property, any desired target sample can be detected simply by introducing with it a reporter which releases a fluorophore when being cut, producing in this way a fluorescent signal that can be measured with a fluorescence detector.
2. Part Collection
This part collection is created with the purpose of helping other future iGEM teams for the guidance in the design of their parts needed to accomplish the full detection system described above. For that, we have created and documented it in a way that it is easy-to-use and user-friendly, as it is as much structured and organized as possible, being all the part pages linked between them in a way that an external user can comfortably replicate our design.
- Categories
As for the collection categories, we propose a basic structure in which each of the gRNA design steps is considered:
| Part Code | Name | Function | Part Collection Category |
|---|---|---|---|
| BBa_K3791000 | Spacer gRNA Ampicillin | Target a specific DNA fragment of a certain ampicillin-resistance gene. | gRNA Spacer |
| BBa_K3791001 | Spacer gRNA Chloramphenicol | Target a specific DNA fragment of a certain chloramphenicol-resistance gene. | gRNA Spacer |
| BBa_K3791002 | Spacer gRNA Erythromycin | Target a specific DNA fragment of a certain erythromycin-resistance gene. | gRNA Spacer |
| BBa_K3791003 | Spacer gRNA Kanamycin | Target a specific DNA fragment of a certain kanamycin-resistance gene. | gRNA Spacer |
| BBa_K3791004 | Spacer gRNA Spectinomycin | Target a specific DNA fragment of a certain spectinomycin-resistance gene. | gRNA Spacer |