Proteins and DNA elements essential for the CRISPR adaptation process in Escherichia coli
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Abstract
The clustered regularly interspaced short palindromic repeats and their associated proteins (CRISPR/Cas) constitute a recently identified prokaryotic defense mechanism against invading nucleic acids. Activity of the CRISPR/Cas system comprises of three steps: (i) insertion of alien DNA sequences into the CRISPR array to prevent future attacks, in a process called 'adaptation', (ii) expression of the relevant proteins, as well as expression and processing of the array, followed by (iii) RNA-mediated interference with the alien nucleic acid. Here we describe a robust assay in Escherichia coli to explore the hitherto least-studied process, adaptation. We identify essential genes and DNA elements in the leader…
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3Topics & keywords
Topics
Keywords
- Biology
- CRISPR
- CRISPR interference
- Trans-activating crRNA
- Nucleic acid
- Genetics
- DNA
- Escherichia coli
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