Difference between revisions of "Part:BBa K3982032"

Revision as of 19:33, 14 October 2021

CODE M Construct C4

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 construct has been assembled using various basic parts to synthesize the customized sgRNA in-vitro for use with Cas14a1 protein.

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
INCOMPATIBLE WITH RFC[10]
Illegal EcoRI site found at 124
Illegal XbaI site found at 148
Illegal PstI site found at 160
12
INCOMPATIBLE WITH RFC[12]
Illegal EcoRI site found at 124
Illegal PstI site found at 160
21
INCOMPATIBLE WITH RFC[21]
Illegal EcoRI site found at 124
Illegal BglII site found at 598
Illegal BamHI site found at 142
23
INCOMPATIBLE WITH RFC[23]
Illegal EcoRI site found at 124
Illegal XbaI site found at 148
Illegal PstI site found at 160
25
INCOMPATIBLE WITH RFC[25]
Illegal EcoRI site found at 124
Illegal XbaI site found at 148
Illegal PstI site found at 160
1000
COMPATIBLE WITH RFC[1000]

@@ Line 3: / Line 3: @@
 <partinfo>BBa_K3982032 short</partinfo>
-CODE M - Construct - C5
+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 construct has been assembled using various basic parts to synthesize the customized sgRNA in-vitro for use with Cas14a1 protein.
-<!-- 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.
 <!-- -->