1 research outputs found
Multiple Chemical Inducible Tal Effectors for Genome Editing and Transcription Activation
Inducible modulation is often required
for precise investigations and manipulations of dynamic biological
processes. Transcription activator-like effectors (TALEs) provide
a powerful tool for targeted gene editing and transcriptional programming.
We designed a series of chemical inducible systems by coupling TALEs
with a mutated human estrogen receptor (ER<sup>T2</sup>), which renders
them 4-hydroxyl-tamoxifen (4-OHT) inducible for access of the genome.
Chemical inducible genome editing was achieved <i>via</i> fusing two tandem ER<sup>T2</sup> domains to customized transcription
activator-like effector nuclease (TALEN), which we termed “Hybrid
Inducible Technology” (HIT-TALEN). Those for transcription
activation were vigorously optimized using multiple construct designs.
Most efficient drug induction for endogenous gene activation was accomplished
with minimal background activity using an optimized inducible TALE
based SunTag system (HIT-TALE-SunTag). The HIT-SunTag system is rapid,
tunable, selective to 4-OHT over an endogenous ligand, and reversible
in drug induced transcriptional activation. Versatile systems developed
in this study can be easily applied for editing and transcriptional
programming of potentially any genomic loci in a tight and effective
chemical inducible fashion