Abstract
Clustered Regularly Interspaced Short Palindromic Repeats (CRISPR)–CRISPR-associated (Cas) systems have revolutionized modern molecular biology. Numerous types of these systems have been discovered to date. Many CRISPR–Cas systems have been used as a backbone for the development of potent research tools, with Cas9 being the most widespread. While most of the utilized systems are DNA-targeting, recently more and more attention is being gained by those that target RNA. Their ability to specifically recognize a given RNA sequence in an easily programmable way makes them ideal candidates for developing new research tools. In this review we summarize current knowledge on CRISPR–Cas systems which have been shown to target RNA molecules, that is type III (Csm/Cmr), type VI (Cas13), and type II (Cas9). We also present a list of available technologies based on these systems.
Highlights
Clustered Regularly Interspaced Short Palindromic Repeats (CRISPR)–CRISPR-associated (Cas) (CRISPR–Cas) systems originate from Prokaryotes, where they serve primarily as a defensive mechanism against mobile genetic elements like phages and plasmids [1]
The Csm6, which is a RNase harnessed by type III CRISPR–Cas systems, was used in the SHERLOCKv2 method of specific RNA detection [68]
It must be noted that RNA-targeting CRISPR–Cas systems are not perfect, and their utilization is facing challenges such as collateral cleavage, off target activity, troublesome delivery of PAMers, significant size of components limiting choice of delivery methods, or even possible immunogenicity that is caused by using proteins of bacterial origin in mammalian cells
Summary
Clustered Regularly Interspaced Short Palindromic Repeats (CRISPR)–CRISPR-associated (Cas) (CRISPR–Cas) systems originate from Prokaryotes, where they serve primarily as a defensive mechanism against mobile genetic elements like phages and plasmids [1]. Discovery of the CRISPR–Cas systems has revolutionized modern molecular biology Due to their unique mechanism, they possess several features highly desirable for potential tools in this field. The CRISPR–Cas has proven to be the most convenient backbone for the development of new tools for research and biotechnology [24,25,26,27] One reason for this is its simplicity, as it requires only one Cas protein for activity. In recent years several new types of CRISPR–Cas systems targeting RNA were discovered, paving the way for the development of new tools for research and biotechnology
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