Abstract
ABSTRACTThe adaptive immune system of prokaryotes, called CRISPR-Cas (clustered regularly interspaced short palindromic repeats and CRISPR-associated genes), results in specific cleavage of invading nucleic acid sequences recognized by the cell’s “memory” of past encounters. Here, we exploited the properties of native CRISPR-Cas systems to program the natural “memorization” process, efficiently generating immunity not only to a bacteriophage or plasmid but to any specifically chosen DNA sequence.
Highlights
The adaptive immune system of prokaryotes, called CRISPR-Cas, results in specific cleavage of invading nucleic acid sequences recognized by the cell’s “memory” of past encounters
The natural process of adaptation in CRISPR-Cas systems, involves the incorporation of short (~30-nucleotide [nt]) spacers, typically derived from foreign genetic elements, into a repeat-spacer array (CRISPR) that forms the memory of the system [1]
When S. thermophilus cells are challenged with virulent phages, approximately one in 106 cells survive by acquisition of a new phage-derived spacer within the CRISPR array [10]; any of the phage protospacers (e.g., 716 in the genome of the S. thermophilus phage 2972 [11]) could form the basis of that immunity
Summary
The adaptive immune system of prokaryotes, called CRISPR-Cas (clustered regularly interspaced short palindromic repeats and CRISPR-associated genes), results in specific cleavage of invading nucleic acid sequences recognized by the cell’s “memory” of past encounters. Tool to bias (and select for) the natural acquisition of specific spacers—in other words, readily programming the native CRISPR arrays. 6% of cells screened after 60 generations had lost the plasmid, 55% of those due to the acquisition of a plasmid-targeting spacer within a CRISPR array.
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