Interference activity of a minimal Type I CRISPR-Cas system from Shewanella putrefaciens

Nucleic Acids Research
Srivatsa DwarakanathLennart Randau

Abstract

Type I CRISPR (Clustered Regularly Interspaced Short Palindromic Repeats)-Cas (CRISPR-associated) systems exist in bacterial and archaeal organisms and provide immunity against foreign DNA. The Cas protein content of the DNA interference complexes (termed Cascade) varies between different CRISPR-Cas subtypes. A minimal variant of the Type I-F system was identified in proteobacterial species including Shewanella putrefaciens CN-32. This variant lacks a large subunit (Csy1), Csy2 and Csy3 and contains two unclassified cas genes. The genome of S. putrefaciens CN-32 contains only five Cas proteins (Cas1, Cas3, Cas6f, Cas1821 and Cas1822) and a single CRISPR array with 81 spacers. RNA-Seq analyses revealed the transcription of this array and the maturation of crRNAs (CRISPR RNAs). Interference assays based on plasmid conjugation demonstrated that this CRISPR-Cas system is active in vivo and that activity is dependent on the recognition of the dinucleotide GG PAM (Protospacer Adjacent Motif) sequence and crRNA abundance. The deletion of cas1821 and cas1822 reduced the cellular crRNA pool. Recombinant Cas1821 was shown to form helical filaments bound to RNA molecules, which suggests its role as the Cascade backbone protein. A Cascade ...Continue Reading

References

Mar 29, 2005·Journal of Molecular Evolution·Francisco J M MojicaElena Soria
Mar 24, 2007·Science·Rodolphe BarrangouPhilippe Horvath
Aug 16, 2008·Science·Stan J J BrounsJohn van der Oost
Jun 13, 2009·Methods in Molecular Biology·Ibtissem GrissaChristine Pourcel
Jan 9, 2010·Science·Philippe Horvath, Rodolphe Barrangou
Mar 30, 2010·Applied and Environmental Microbiology·Jürgen LassakKai M Thormann
Jul 14, 2010·Molecular Microbiology·Ksenia PougachKonstantin Severinov
Sep 11, 2010·Science·Rachel E HaurwitzJennifer A Doudna
Sep 28, 2010·Methods in Cell Biology·Reinhard RachelGerhard Wanner
Oct 5, 2010·RNA·Jason CarteMichael P Terns
Oct 28, 2010·BioEssays : News and Reviews in Molecular, Cellular and Developmental Biology·Adi Stern, Rotem Sorek
Nov 5, 2010·Nature·Josiane E GarneauSylvain Moineau
Mar 15, 2011·Journal of Bacteriology·Kyle C Cady, George A O'Toole
Apr 2, 2011·Nature·Elitza DeltchevaEmmanuelle Charpentier
Apr 5, 2011·Nature Structural & Molecular Biology·Matthijs M JoreStan J J Brouns
May 4, 2011·Proceedings of the National Academy of Sciences of the United States of America·Blake WiedenheftJennifer A Doudna
May 10, 2011·Nature Reviews. Microbiology·Kira S MakarovaEugene V Koonin
May 17, 2011·Nature Structural & Molecular Biology·Emily M GesnerAndrew M Macmillan
Apr 24, 2012·Molecular Cell·Dipali G SashitalJennifer A Doudna
Jun 30, 2012·Science·Martin JinekEmmanuelle Charpentier
Nov 14, 2012·Annual Review of Genetics·Edze R WestraJohn van der Oost
Mar 16, 2013·RNA Biology·Ambarish BiswasChris M Brown
Apr 9, 2013·RNA Biology·Krzysztof ChylinskiEmmanuelle Charpentier
Apr 27, 2013·RNA Biology·Ekaterina SavitskayaKonstantin Severinov
Jun 29, 2013·The Biochemical Journal·Judith ReeksMalcolm F White
Oct 15, 2013·Molecular Cell·Christophe RouillonMalcolm F White
Oct 24, 2013·Nucleic Acids Research·Ole NiewoehnerJennifer A Doudna
Nov 22, 2013·Biochemical Society Transactions·Judith Zoephel, Lennart Randau
Jan 15, 2014·PLoS Genetics·Nina SestoPascale Cossart
Feb 7, 2014·Nucleic Acids Research·André PlagensLennart Randau
Apr 22, 2014·Proceedings of the National Academy of Sciences of the United States of America·Megan L HochstrasserJennifer A Doudna

❮ Previous
Next ❯

Citations

May 25, 2016·Nucleic Acids Research·Daniel GleditzschLennart Randau
Jun 13, 2020·Frontiers in Molecular Biosciences·Hanna Müller-EsparzaLennart Randau
Dec 8, 2019·Nature Communications·Hiroyuki MorisakaTomoji Mashimo
Jun 8, 2021·Frontiers in Microbiology·Hannah N TaylorRyan N Jackson
Sep 7, 2021·International Microbiology : the Official Journal of the Spanish Society for Microbiology·Noemí M GuzmánFrancisco J M Mojica

❮ Previous
Next ❯

Methods Mentioned

BETA
PCR
RNA-Seq
gene-knockout
electrophoresis
size exclusion chromatography
electron microscopy

Software Mentioned

CRISPRtarget
Agilent QuickChange Primer Design Tool
Cascade

Related Concepts

Related Feeds

CRISPR (general)

Clustered regularly interspaced short palindromic repeats (CRISPR) are DNA sequences in the genome that are recognized and cleaved by CRISPR-associated proteins (Cas). CRISPR-Cas system enables the editing of genes to create or correct mutations. Discover the latest research on CRISPR here.

CRISPR Genome Editing & Therapy

CRISPR-Cas system enables the editing of genes to create or correct mutations. This feed focuses on the application of this system for gene editing and therapy in human diseases.

CRISPR & Staphylococcus

CRISPR-Cas system enables the editing of genes to create or correct mutations. Staphylococci are associated with life-threatening infections in hospitals, as well as the community. Here is the latest research on how CRISPR-Cas system can be used for treatment of Staphylococcal infections.

CRISPR for Genome Editing

Genome editing technologies enable the editing of genes to create or correct mutations. Clustered regularly interspaced short palindromic repeats (CRISPR) are DNA sequences in the genome that are recognized and cleaved by CRISPR-associated proteins (Cas). Here is the latest research on the use of CRISPR-Cas system in gene editing.

CRISPR Ribonucleases Deactivation

CRISPR-Cas system enables the editing of genes to create or correct mutations. This feed focuses on mechanisms that underlie deactivation of CRISPR ribonucleases. Here is the latest research.