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Cloning vector pUC19

ABSTRACT: Sequencing of reaction products from protospacer integration by Cas1-Cas2

PROVIDER: PRJNA271290 | ENA |

REPOSITORIES: ENA

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			Action	DRS
	SRR1736304.fastq.gz	Fastqsanger.gz
	SRR1736305.fastq.gz	Fastqsanger.gz
	SRR1736306.fastq.gz	Fastqsanger.gz
	SRR1736307.fastq.gz	Fastqsanger.gz

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Sequencing of reaction products from protospacer integration by Cas1-Cas2

Project description:We report the sites of protospacer integration into plasmids by Cas1-Cas2. The goal of this study is to determine the role of sequence elements on protospacer integration. Methods: Cas1-Cas2 integration products were fragmented, end-repaired, adapter ligated, PCR amplified, and analyzed by Illumina sequencing. Protospacer-containing reads were selected by Cutadapt and mapped to the plasmid with Bowtie.

2015-01-30 | E-GEOD-64552 | biostudies-arrayexpress

Cloning vector pUC19

Project description:Protospacer integration into plasmid DNA by Cas1–Cas2

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Sequencing of reaction products from protospacer integration by Cas1-Cas2

Project description:We report the sites of protospacer integration into plasmids by Cas1-Cas2. The goal of this study is to determine the role of sequence elements on protospacer integration.

2015-01-30 | GSE64552 | GEO

Protospacer integration into plasmid DNA by Cas1–Cas2

Project description:We report the integration site specificity of Cas1–Cas2 into target plasmid DNA encoding a CRISPR locus. The goal of this study is to determine the effect of IHF on target site selection of Cas1–Cas2.

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Project description:Key to CRISPR-Cas adaptive immunity is maintaining an ongoing record of invading nucleic acids that are encountered, a process carried out by the Cas1-Cas2 complex that integrates short segments of foreign genetic material (spacers) into the CRISPR locus. It is hypothesized that Cas1 evolved from casposases, a novel class of transposases. We show here that casposase integration in vitro recapitulates several properties of CRISPR-Cas integrases. The X-ray structure of Methanosarcina mazei casposase bound to DNA representing the product of integration reveals a tetramer with target DNA bound snugly between two dimers in which single-stranded casposon end binding resembles that of spacer 3'-overhangs. The differences between transposase and CRISPR-Cas integrase are largely architectural, and it appears that evolutionary change involved changes in protein-protein interactions to favor Cas2 binding over tetramerization and the separation of Cas1 dimers. This, in turn, led to preferred integration of single spacers over two transposon ends.

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