Directed evolution, bioorthogonal chemistry, click chemistry, PROTACs, unnatural amino acids.
Systems And Chemical Biology Methods
Directed evolution
Iterative mutagenesis + selection/screening to engineer proteins (enzymes, binders) with new functions. Arnold Nobel 2018. Error-prone PCR,…
Click chemistry
Selective bio-orthogonal reactions for in-vivo labelling. CuAAC (Cu-catalysed azide-alkyne cycloaddition) + SPAAC (strain-promoted,…
Bioorthogonal chemistry
Chemistry that proceeds inside living systems without interfering with native biochemistry. Staudinger ligation, tetrazine ligation, SPAAC.…
PROTAC (proteolysis-targeting chimera)
Bifunctional small molecule linking target protein to E3 ubiquitin ligase → degradation via UPS. Catalytic, event-driven pharmacology —…
Unnatural amino acid (UAA) incorporation
Site-specific insertion of non-canonical AAs via orthogonal aaRS/tRNA pair and reassigned stop codon (amber). Enables photocrosslinking,…
Activity-based probe
Small-molecule covalent probe that labels only active/unliganded enzymes. Reports enzyme activity in situ. Cravatt: serine hydrolase ABPs…
Chemical proteomics
MS-based identification of protein targets of small molecules. Affinity pull-downs, thermal proteome profiling (TPP), drug affinity…
CRISPR genetic screen
Genome-wide knockout/activation libraries (GeCKO, Brunello) + phenotypic readout. Pooled or arrayed. Identifies synthetic lethal partners,…
High-throughput screening
Automated assay of ~10⁵-10⁶ compounds against a biological target. Plate-based fluorescence/luminescence. Hit-to-lead to clinical candidate…
Optogenetics
Light-gated ion channels (channelrhodopsin, halorhodopsin) expressed in defined neurons. Millisecond control of activity in vivo.…