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Unbiased profiling of CRISPR RNA-guided transposition products by long-read sequencing

Mob DNA. 2021-06; 
Phuc Leo H Vo, Christopher Acree, Melissa L Smith, Samuel H Sternberg
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Gene Synthesis … TnsAB fusions producing almost no detectable cointegrates across a population of cells (Fig … for applications of RNA-guided DNA integration technology in heterologous cell types … 09/160 genome (NCBI accession LR721750.1), synthesized as fragments (GenScript), and cloned … Get A Quote

摘要

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 e... More

关键词

CAST, CRISPR-Cas, RNA-guided transposase, SMRT-seq, Tn7, Transposition
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