Processing and integration of functionally oriented prespacers in the Escherichia coli CRISPR system depends on bacterial host exonucleases
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TL;DR: It is shown that the bacterial 3′–5′ exonucleases DnaQ and ExoT can trim long 3′ overhangs of prespacers and promote integration in the correct orientation, and found that trimming by these exon nucleases results in an asymmetric intermediate, because Cas1-Cas2 protects the PAM sequence, which helps to define spacer orientation.
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About: This article is published in Journal of Biological Chemistry. The article was published on 13 Mar 2020. and is currently open access. The article focuses on the topics: Protospacer adjacent motif & dnaQ.
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Citations
Molecular Mechanisms of CRISPR-Cas Immunity in Bacteria.
TL;DR: This work describes the many different molecular mechanisms of CRISPR targeting and how they are interconnected with the immunization phase through a third phase of theCRISPR-Cas immune response: primed spacer acquisition.
150
Structural biology of CRISPR–Cas immunity and genome editing enzymes
TL;DR: The adaptive and interference aspects of CRISPR–Cas function as well as open questions about the molecular mechanisms responsible for genome targeting are explored; structural insights reflect close evolutionary links between CRISpr–Cas systems and mobile genetic elements.
125
Recording gene expression order in DNA by CRISPR addition of retron barcodes
Santi Bhattarai-Kline,Sierra K Lear,C. Fishman,Santiago Cuesta López,Elana R. Lockshin,Max G. Schubert,Jeff Nivala,George M. Church,Seth L. Shipman +8 more
TL;DR: In this article , a molecular system for making time-ordered recordings of transcriptional events into living genomes is presented, which is based on engineered RNA barcodes, based on prokaryotic retrons, that are reverse transcribed into DNA and integrated into the genome using the CRISPR-Cas system.
57
Mechanism for Cas4-assisted directional spacer acquisition in CRISPR–Cas
Chunyi Hu,Cristóbal Almendros,Cristóbal Almendros,Ki Hyun Nam,Ana Rita Costa,Ana Rita Costa,Jochem N.A. Vink,Jochem N.A. Vink,Anna C. Haagsma,Anna C. Haagsma,Saket R. Bagde,Stan J. J. Brouns,Stan J. J. Brouns,Ailong Ke +13 more
TL;DR: In this article, a high-resolution mechanistic explanation for the Cas4-assisted PAM selection, spacer biogenesis and directional integration by type I-G CRISPR in Geobacter sulfurreducens was provided.
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Creating memories: molecular mechanisms of CRISPR adaptation.
Hayun Lee,Dipali G. Sashital +1 more
TL;DR: A review of recent advances in CRISPR adaptation can be found in this article , with a focus on how functional spacers are generated and incorporated through many widespread, but type-specific, mechanisms.
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