Fig. 4: Specificity of target recognition by D-R pAgos. | Nature Communications

Fig. 4: Specificity of target recognition by D-R pAgos.

From: Programmable RNA targeting by bacterial Argonaute nucleases with unconventional guide binding and cleavage specificity

Fig. 4

a Binding of target RNA and ssDNA of the same sequence by PliAgo, PnyAgo and CbAgo loaded with cognate guide DNA. Means and standard deviations from 3 independent experiments are shown. b Scheme of RNA/DNA targets containing a single dG/rG residue with differently positioned DNA guides (see Supplementary Table 1 for oligonucleotide sequences). Positions of the functional regions in guide DNA are indicated (seed, orange; central, magenta; 3’-supplementary region, ochre). c Effects of a single deoxyribonucleotide (dG) in the RNA target (top) and a single ribonucleotide (rG) in the DNA target (bottom) on target cleavage by PliAgo in comparison with control RNA and DNA targets. The reaction was performed at 37 °C for 30 min in the case of target RNA and for 3 h in the case of target DNA. Means from 2 independent experiments are shown. Target positions around the cleavage site are shown with dotted boxes. d Kinetics of target RNA (left) or target DNA (right) cleavage by PliAgo with containing ribonucleotide (rG) or deoxyribonucleotide (dG) residues at the position 10’. Representative gels from two independent experiments are shown.

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