Figure 1. Gut native E. coli are genetically tractable and can serve as a chassis for transgene delivery.

(A) Experimental strategy of engineered native bacteria.

(B) Optimal characteristics of a chassis for transgene delivery for a potential LBT.

(C) Original isolate (EcAZ-1), GFP-producing strain (EcAZ-2), and GFP- and BSH-producing strain (EcAZ-2^BSH+) plated on LB containing TDCA. Precipitate around EcAZ-2^BSH+ is the result of TDCA deconjugated by the bacteria to DCA and qualitatively indicates enzyme functionality.

(D) Growth curve of EcAZ-2 compared with EcAZ-1, EcAZ-2^BSH+ compared with EcAZ-1, and EcAZ-2^IL10+ compared with EcAZ-2. The line represents an average of three measurements per strain.

(E) Proteomic analysis shows increased BSH protein expression in EcAZ-2^BSH+ compared with EcAZ-1 and EcAZ-2 (n = 4).

(F) Log₂ ratio of DCA to TDCA in a timed enzymatic assay (n = 3; see Figure S1G for raw values).

(G) IL-10 levels detected with ELISA from cell lysates (n = 6).

The marker covers some error bars in (F) and (G). Given the low number of samples for (E) and (F), we used a Student’s t test after the confirmation of normality with Q-Q plot. We used a Mann-Whitney U test for (G). (A) and (B) were created with BioRender.com.

See also Figure S1.