Expanding and understanding the CRISPR toolbox for Bacillus subtilis with MAD7 and dMAD7

Feb 21, 2020Biotechnology and bioengineering

Using MAD7 and dMAD7 to improve gene editing tools for Bacillus subtilis

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Abstract

Editing rates of 93% and 100% were established using the MAD7 nuclease for gene editing in Bacillus subtilis.

  • MAD7 is a newly available CRISPR nuclease that can be used without licensing fees for industrial research and development.
  • The first catalytically inactive variant of MAD7 (dMAD7) was engineered, demonstrating its potential for gene regulation.
  • dMAD7 achieved up to 71.3% reduction in gene expression at single and multiple target sites in B. subtilis.
  • The research confirmed that the DNA double-strand breaks induced by the nuclease facilitate selection during gene editing.

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