Fig. 2
From: Unbiased profiling of CRISPR RNA-guided transposition products by long-read sequencing
RNA-guided transposition with Tn7-like CRISPR-transposons encoding TnsAB fusion proteins. a Genetic architecture of A. wodanis transposon harboring a natural tnsAB fusion gene, with pfam domains for the TnsAB protein shown below. b PCR analysis and Sanger sequencing confirm RNA-guided DNA integration with A. wodanis CRISPR-Tn. NT, non-targeting. c qPCR analysis showing integration activity with the V. cholerae CRISPR-Tn, for a pEffector encoding either TnsA and TnsB (A+B) or a fusion TnsAB polypeptide (ABf), with WT or D90A mutation. Data are shown as mean ± s.d. for 3 biological replicates. d Representative SMRT-seq reads from the V. cholerae CRISPR-Tn with fused TnsAB, showing hallmarks of simple insertion products (top); SMRT-seq reads from the same system with D90A-TnsAB are shown at bottom, showing hallmarks of cointegrate products. e Population-level quantification of simple insertion and cointegrate transposition products from the V. cholerae CRISPR-Tn with TnsAB fusion, for either the WT system or a D90A-TnsAB mutant. Data represent mean ± s.d. for 3 biological replicates; n denotes the total number of CCS reads. f Specificity of RNA-guided transposition determined from SMRT-seq data, for the V. cholerae CRISPR-Tn with TnsAB fusion. Data are shown as mean for 3 biological replicates; n denotes the total number of CCS reads