Part:BBa_K5096046

inhA target

The sequence encodes the complete inhA sgRNA target, which can be utilized to test the inhA sgRNA’s region. sgRNA70 targets the inhA region of the M. tuberculosis genome. The binding sequence of sgRNA70 is derived from the inhA gene, chosen for its low mutation rate and essential role in the pathogenicity and survival of M. tuberculosis. The inhA gene encodes NADH-dependent enoyl-acyl carrier protein reductase, a crucial enzyme in mycolic acid synthesis, a vital component of the bacterial cell envelope of M. tuberculosis (Marrakchi et al., 2014). By inhibiting mycolic acid synthesis through the CRISPRi system, sgRNA70 seeks to destabilize the bacterial cell wall, thereby weakening the pathogen.

Lambert iGEM also utilized MATLAB to perform precise modeling of gene expression of inhA, a gene linked to the pathogenicity of M. tuberculosis (see CRISPRi M. tb POC for more about inhA). We created a complete set of ODEs, and then used Simbiology’s Model Analyzer to simulate the concentration of GFP over 20 hours with 1 nanomole of the inhA DNA initially. As we are producing GFP protein, the graph will increase at first, and then around four hours, the graph will plateau. <figure>

   <img src="" width="50%">

</figure>

Sequence and Features

Assembly Compatibility:

• 10
  COMPATIBLE WITH RFC[10]
• 12
  INCOMPATIBLE WITH RFC[12]

  Illegal NheI site found at 1030
  
• 21
  COMPATIBLE WITH RFC[21]
• 23
  COMPATIBLE WITH RFC[23]
• 25
  COMPATIBLE WITH RFC[25]
• 1000
  COMPATIBLE WITH RFC[1000]