Abstract
Cas3 has essential functions in CRISPR immunity but its other activities and roles, in vitro and in cells, are less widely known. We offer a concise review of the latest understanding and questions arising from studies of Cas3 mechanism during CRISPR immunity, and highlight recent attempts at using Cas3 for genetic editing. We then spotlight involvement of Cas3 in other aspects of cell biology, for which understanding is lacking—these focus on CRISPR systems as regulators of cellular processes in addition to defense against mobile genetic elements.
Highlights
Cas3 has essential functions in CRISPR immunity but its other activities and roles, in vitro and in cells, are less widely known
Cas3 is an ATP-dependent single-strand DNA translocase/helicase that in many CRISPR systems is fused to a HD-nuclease domain
Cas3 was first highlighted in 2002 during in silico analyses of prokaryotic genomes [4,5] that identified its superfamily-2 helicase motifs [4], and its association with other proposed nucleic acid processing enzymes located alongside repeat DNA sequences [5]
Summary
We review original research that describes the structure and function of prokaryotic. Cas was first highlighted in 2002 during in silico analyses of prokaryotic genomes [4,5] that identified its superfamily-2 helicase motifs (as ‘COG1203’) [4], and its association with other proposed nucleic acid processing enzymes located alongside repeat DNA sequences [5]. This created the term ‘CRISPR’, for the DNA repeats, and the protein COG1203 became ‘Cas3’, a CRISPR-associated protein [5]—the discovery and establishment of CRISPR biology and biotechnology is described in [6]. We highlight experimental observations that indicate potential roles for Cas in RNA and biofilm formation, and usefulness in genetic editing reactions
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