Instruct: Identify the main category of Biorxiv papers based on the titles and abstracts
Query: Unbiased profiling of CRISPR RNA-guided transposition products by long-read sequencing Bacterial transposons propagate through either non-replicative (cut-and-paste) or replicative (copy-and-paste) pathways, depending on how the mobile element is excised from its donor source. In the well-characterized E. coli transposon Tn7, a heteromeric TnsA-TnsB transposase directs cut-and-paste transposition by cleaving both strands at each transposon end during the excision step. Whether a similar pathway is involved for RNA-guided transposons, in which CRISPR-Cas systems confer DNA target specificity, has not been determined. Here, we apply long-read, population-based whole-genome sequencing (WGS) to unambiguously resolve transposition products for two evolutionarily distinct transposon types that employ either Cascade or Cas12k for RNA-guided DNA integration. Our results show that RNA-guided transposon systems lacking functional TnsA primarily undergo copy-and-paste transposition, generating cointegrate products that comprise duplicated transposon copies and insertion of the vector backbone. Finally, we report natural and engineered transposon variants encoding a TnsAB fusion protein, revealing a novel strategy for achieving RNA-guided transposition with fewer molecular components.
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The categories of this paper are: biochemistry
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The categories of this paper are: genetics
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The categories of this paper are: and
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The categories of this paper are: epidemiology
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The categories of this paper are: bioinformatics
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The categories of this paper are: synthetic
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The categories of this paper are: genomics
3
The categories of this paper are: evolutionary
21
The categories of this paper are: toxicology
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The categories of this paper are: cell
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The categories of this paper are: education
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The categories of this paper are: animal
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The categories of this paper are: pathology
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The categories of this paper are: bioengineering
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The categories of this paper are: behavior
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The categories of this paper are: physiology