Building a Genetic Switch in Bacteria to Reduce Protein Production

The Tl-CRISPRi system enables programmable gene knockdown in bacteria.

• The system utilizes the RNA-binding activity of CRISPR-dCas13 to inhibit mRNA translation.
• Specific spacers complementary to the translation initiation region of mRNA block the ribosome.
• Methods for constructing the Tl-CRISPRi genetic circuit are detailed, including spacer design.
• Mutations in the guide RNA handle may allow for adjustable levels of gene knockdown.
• The approach aims for precise control over gene expression in bacterial cells.

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