RT Journal Article
SR Electronic
T1 Connecting Boolean Integrase Logic gates to a novel alkane control signal via engineered level matching
JF bioRxiv
FD Cold Spring Harbor Laboratory
SP 031542
DO 10.1101/031542
A1 Thomas Folliard
A1 Jerome Bonnet
A1 John Ward
A1 Frank Baganz
A1 Chris Grant
A1 Drew Endy
YR 2015
UL http://biorxiv.org/content/early/2015/11/12/031542.abstract
AB Boolean Integrase Logic gates use phage integrases that respond to transcriptional signals to implement logical functions within living cells via DNA recombination. Control of biological systems using these logic gates has many applications, including biomanufacturing, healthcare, or environmental remediation. Gates are highly sensitive to background transcriptional noise producing unwanted integrase expression and uncontrolled, permanent gate switching. Consequently, connecting gates to novel control signals requires time-consuming directed evolution of regions regulating integrase expression. Here we present an approach in which the activity of an alkane biosensor is tuned to match levels of existing integrase control signals. By adjusting AlkS expression, we tuned the transcriptional output of Palkb to match the output of existing integrase controllers. We successfully connected Palkb to two integrases with different transcriptional control requirements and demonstrated the original logic function was conserved. Our method complements directed evolution approaches to connect Boolean integrate logic gates to novel transcriptional sources and will ultimately facilitate the systematic in silico design of gates responding to various control signals.