Difference between revisions of "Part:BBa K3982039"

Latest revision as of 15:37, 21 October 2021

CODE M Construct S3

This construct has been used in Project CODE M by Team IISER_Berhampur 2021. The aim of this project is to combat Multidrug-Resistant Tuberculosis (MDR-TB) with our diagnostic kit CODE M using mobile phone microscopy. Mutant strains of Mycobacterium tuberculosis, the causative agent of MDR-TB (which confer drug resistance towards two main drugs for TB cure - isoniazid and rifampicin) are detected through high-fidelity SNP detection using CRISPR/Cas technology. Here we used Cas14a1 (also called as Un1Cas12f1) as the RNA guided nuclease with our set of customised CODE M sgRNAs.

This is a standard construct has been assembled using various new basic parts as well as existing parts to synthesize the customized sgRNA in-vitro for use with Cas14a1 protein. The purpose of this construct is to test the expression of our CODE M sgRNAs with some common regulatory parts.

Usage and Biology

CRISPR/Cas technology has been derived from the CRISPR/Cas systems present in archaea and bacteria. They function as an adaptive immune system against invading foreign DNA and RNA. This technology has grown in the past several years with a wide range of applications. There are different types of Cas proteins such as Cas9, Cas12, Cas13 as well as Cas14. All of them are RNA guided nucleases - with a crRNA and tracrRNA or a sgRNA.

sgRNAs have significantly evolved from the natural guide RNAs which consisted of two parts - crRNA and tracrRNA. The optimized structure of sgRNA makes it highly efficient for genome editing comparable to that of spCas9 guide RNAs.

Sequence and Features

Assembly Compatibility:

10
COMPATIBLE WITH RFC[10]
12
COMPATIBLE WITH RFC[12]
21
INCOMPATIBLE WITH RFC[21]
Illegal BglII site found at 406
23
COMPATIBLE WITH RFC[23]
25
COMPATIBLE WITH RFC[25]
1000
COMPATIBLE WITH RFC[1000]

@@ Line 3: / Line 3: @@
 <partinfo>BBa_K3982039 short</partinfo>
-TC2
+This construct has been used in Project CODE M by [https://2021.igem.org/Team:IISER_Berhampur Team IISER_Berhampur 2021]. The aim of this project is to combat Multidrug-Resistant Tuberculosis (MDR-TB) with our diagnostic kit CODE M using mobile phone microscopy. Mutant strains of Mycobacterium tuberculosis, the causative agent of MDR-TB (which confer drug resistance towards two main drugs for TB cure - isoniazid and rifampicin) are detected through high-fidelity SNP detection using CRISPR/Cas technology. Here we used Cas14a1 (also called as Un1Cas12f1) as the RNA guided nuclease with our set of customised CODE M sgRNAs.
+This is a standard construct has been assembled using various new basic parts as well as existing parts to synthesize the customized sgRNA in-vitro for use with Cas14a1 protein. The purpose of this construct is to test the expression of our CODE M sgRNAs with some common regulatory parts.
-<!-- Add more about the biology of this part here
 ===Usage and Biology===
+CRISPR/Cas technology has been derived from the CRISPR/Cas systems present in archaea and bacteria. They function as an adaptive immune system against invading foreign DNA and RNA. This technology has grown in the past several years with a wide range of applications. There are different types of Cas proteins such as Cas9, Cas12, Cas13 as well as Cas14. All of them are RNA guided nucleases - with a crRNA and tracrRNA or a sgRNA.
+sgRNAs have significantly evolved from the natural guide RNAs which consisted of two parts - crRNA and tracrRNA. The optimized structure of sgRNA makes it highly efficient for genome editing comparable to that of spCas9 guide RNAs.
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