Fig. 2: Molecular design and operational principles of logic gate-responsive DNA receptors. | Nature Communications

Fig. 2: Molecular design and operational principles of logic gate-responsive DNA receptors.

From: Synthetic transmembrane DNA receptors enable engineered sensing and actuation

Fig. 2: Molecular design and operational principles of logic gate-responsive DNA receptors.The alternative text for this image may have been generated using AI.

a The design of AND logic gate-responsive DNA receptors (A-Seq1 and A-Seq2) on the cell surface for precise cell identification. Two recognition aptamers (A-apt1 and A-apt2) used for specifically targeting MUC 1 and Met, which are blocked by partially complementary DNAs (L-A1 and L-A2), two C12 spacers used for anchoring live cells bound with aptamers, and intracellular parts of A-Seq1 and A-Seq2 (A-C1 and A-C2) conjugated with dyes for monitoring DNA dimerization. b The design of OR logic gate-responsive DNA receptors (O-Seq1 and O-Seq2) on the cell surface for precise cell identification. Two recognition aptamers (O-apt1 and O-apt2) used for specifically targeting MUC 1 and Met, which are blocked by partially complementary DNAs (L-O1 and L-O2), two C12 spacers used for anchoring live cells bound with aptamers, and intracellular parts of O-Seq1 and O-Seq2 (O-C1 and O-C2) conjugated with dyes for monitoring DNA dimerization. c, d CLSM images of four cell lines incubated with AND and OR logic gate-responsive signal receptors at 37 C for 2 h. Step 1: the recognition between AptMUC 1 and MUC 1 after adding A-Seq1/O-Seq1; step 2: the recognition between AptMet and Met after adding A-Seq2/O-Seq2 and the dimerization between A-Seq1/O-Seq1 and A-Seq2/O-Seq1. Scale bar: 10 μm. e, f FL intensity comparison histogram of different fluorochromes per cell in (c, d). g, h PAGE analysis of the stepwise assembly of logic gate-responsive signal receptors. Lane 1 to 7 in AND logic gate: ladder (lane 1), A-Seq1 (lane 2), A-Seq2 (lane 3), L-A1 (lane 4), L-A2 (lane 5), a mixture of A-Seq1 and L-A1 (lane 6), a mixture of A-Seq2 and L-A2 (lane 7). Lane 1 to 7 in OR logic gate: ladder (lane 1), O-Seq1 (lane 2), O-Seq2 (lane 3), L-O1 (lane 4), L-O2 (lane 5), a mixture of O-Seq1 and L-O1 (lane 6), a mixture of O-Seq2 and L-O2 (lane 7). Experiments were repeated three times independently with similar results. Error bars in (e, f) represent the standard deviation of three independent experimental repeats, and the measure of the center represents their corresponding mean value. Source data are provided as a Source Data file.

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