...provided by the Rinehart lab. Steinfeld et al., ACS Chem Biol 2014 May 16;9(5):1104-12. Heinemann et... your own designer bacteria. St-Pierre et al., ACS Synth Biol 2013 Sep 20;2(9):537-41. DREADD-based... One. 2011. Feb 18;6(2):e16765. Engler et al., ACS Synth Biol 2014. Feb 5 (Web); DOI: 10.1021/sb4001504...endogenous protein of interest and co-expressing its replacement recombinant protein with a FKBP domain and a ...tyrosine kinases (RTKs) are a large class of cell-surface receptors which play a critical role in development...dimerize on exposure to light, bringing the intracellular kinase domains into contact and initiating signaling...optimal scaffold can depend on the length of the spacer region between the target sequences. These updates...

...Wingler L, Pascolutti R, Paulo JA, Gygi SP, Kruse AC (2017) Multidimensional Tracking of GPCR Signaling...is initiated when a ligand binds to the extracellular surface of the GPCR. This results in a conformational...induce or inhibit intracellular signaling cascades as a response to the extracellular stimuli. Ligand dissociation...coupled with heterotrimeric G proteins on the intracellular side of the cell membrane. Heterotrimeric G ...relatively insensitive to their endogenous ligand, acetylcholine, and are only activated upon binding of CNO ...

...plasmids from Aranda-Díaz et al. Aranda-Díaz, et al. ACS Synth Biol. 2017. Pubmed PMID: 27930885. ...or noise, in reporter expression. To do so, one places reporter expression under the control of two separate...

...retain some ability to induce Cas9 to make edits (AC, WLC, and JQ, unpublished results). An additional...that is complementary to the target site, or protospacer, in the genome. Native Cas9 has two catalytic...which cleaves one strand of DNA upon binding the protospacer. The resulting double strand break (DSB) stimulates...

... the decaffeination plasmid here. Quandt et al. ACS Synth Biol. 2013. PubMed PMID: 23654268. FLINC...OptoSTIM1), caused two-fold faster changes in intracellular Ca2+ levels than the original version containing...substrate, inducing a conformational change that placesTagRFP-T and Dronpa in close proximity. By monitoring...

...editing toolbox.  References Gaudelli NM, Komor AC, Rees HA, Packer MS, Badran AH, Bryson DI, Liu DR...in the genomic DNA to contain a specific protospacer adjacent motif (PAM) made up of 5′-NGG-3′ (“N” can...eighth round of ABE evolution and the “x” is the placeholder for the number that identifies the mutation in...to ABE7.10. Depending on the position in the protospacer, the editing capacity of the ABE8s ranged from...To expand the targeting scope of ABE8, the team replaced S. pyogenes Cas9 in the ABE complex with PAM-variant...

...Jones, Nicola J. Patron, and Sylvestre Marillonnet. ACS Synth. Biol., Just Accepted Manuscript, DOI: 10.1021...

...Receptors Exclusively Activated by Designer Drugs. ACS Chemical Neuroscience 6:476–484 . https://doi.org...nonnative ligands. However, they retain their intracellular interactions and still set off the same pathways...

...Integration Using CRISPR-Associated Transposases. ACS Synth Biol 9:1998–2008 . https://doi.org/10.1021/...top):  A CRISPR RNA (crRNA) comprising a 32 nt spacer sequence specifying the target site. Four proteins...INTEGRATE using a single plasmid containing a multi-spacer CRISPR array. Workflow and technical considerations...INTEGRATE system follows a simple workflow: Clone spacer(s) into the CRISPR array Clone desired DNA cargo...

...Ronald T. Raines. "Bright ideas for chemical biology."ACS chemical biology 3.3 (2008): 142-155. PubMed PMID...from ensemble imaging to single-molecule imaging places a greater burden on the photon budget, which determines...

...H3 me1, me3, and ac – denoting mono-methylation, tri-methylation, and aceylation, respectively (these...a genomic region that has mono-methylated and acetylated histones, but not tri-methylated histones, functions...denote enhancers as follows: (mono_methylation + acetylation) - tri_methylation In the next section we’ll ...interests span machine learning, proteomics, and user interfaces for math.     Additional Resources Beyond the...

...References and resources References Bertiaux E, Balestra AC, Bournonville L, et al (2021) Expansion microscopy...?  ExM provides a straightforward answer to the space problem. Instead of imaging smaller, what if we ...

...Integration Using CRISPR-Associated Transposases. ACS Synth Biol 9:1998–2008. https://doi.org/10.1021/acssynbio...the cell. The edit in the chromosome can now take place. After editing, pTargetF can be cured from the cell... E. coli with the donor DNA for editing to take place. Then plasmids are cured from the cell. Steps 1-...involved the following changes. In pCas9, we 1) replaced the promoter of gRNA-pMB1 with PrhaB, 2) changed...counterselection to yield pEcCas. In pTargetF, we replaced the original 20 bp specific sequences on pTargetF...serves as a counterselection marker and will be replaced by the sgRNA targeting the genomic site of interest... clearance. It is more convenient to update the spacer(20nt) on pEcgRNA plasmid as only two 24 nt oligos...

...Orthogonal Ribosome-Controlled Fluorescent Reporter."ACS chemical biology 11.1 (2015): 31-37. pCri system...

...Systems: The Rho-Family GTPase Signaling Toolbox. ACS Synthetic Biology, 11(1), 515–521. https://doi.org...and study a protein’s function at a very specific place and time. Future studies with these CIB1-Rabs in...

...Pillai-Kastoori L, Heaton S, Shiflett SD, Roberts AC, Solache A, Schutz-Geschwender AR (2019) Antibody...sample migrate through the stacking gel at the same pace and enter the resolving gel at the same time where...post-translational modifications such as phosphorylation, acetylation, methylation and ubiquitination. For example,...

...through Structure-Guided and Genome Mining Strategies. ACS Sens. 2022 May 27;7(5):1336-1346. doi: 10.1021/acssensors...sensor Ace2N and additional GCaMP6 constructs: pAAV-CAG-dLight1.2 (AAV1) pGP-AAV-syn-FLEX-Ace2N-4AA-mNeon-ST...fluorescence unit fused with TRansmembrane region-to-IntrAcellular loop linkers” (MTRIA for short), where a fluorescent...inserting a circularly-permuted GFP into an intracellular loop of various G-protein-coupled receptors....fluorescent sensor for real-time measurement of extracellular oxytocin dynamics in the brain. Nat Methods....www.addgene.org/browse/article/28229029/ Lu K, et al. Intracellular Heat Transfer and Thermal Property Revealed ...

... Light-Inducible Gene Editing and Transcription. ACS Chem Biol 13:443–448 . https://doi.org/10.1021/acschembio...With their light-activated CRISPR/Cas9 effector (LACE) system, the Gersbach lab utilized a similar strategy...optimal fusion protein combination. The optimized LACE system consisted of: 1) CIBN-dCas9-CIBN, where CIBN...Far-red light, however, can penetrate 5 mm past the surface of skin and is less phototoxic. Using the FAST ...Activation of gene transcription Polstein et al., 2015 LACE CRY2 and CIB1 from A. thaliana dCas9 Activation ...

...or repression. The TCS light sensors described in ACS Synthetic Biology (2014) represent the newest versions...Photobiology: Optogenetics for E. coli. Schmidl, et al. ACS Synth Biol. 2014 Nov 21;3(11):820-31. Inncredible...compartment (e.g. the plasma membrane, the extracellular space, or the lysosomes) can take many paths. Understanding...that is quick, versatile, and robust. Dubbed the LACE system for light-activated CRISPR-Cas9 effector,...the CRISPR system. The ease and flexibility of the LACE technology makes this system widely accessible for...thereby creating a single-molecule reporter of intracellular processes. Multimerization via the SunTag system...combination of all 11 att pairs in series with unique spacer sequences (pMemoryArray) gives a DNA sequence capable...

...plasmids shipped as agar stabs. Iverson, et al. ACS Synth Bio. 2015. PubMed PMID: 26479688. RAS Pathway...modifiers like DNA methyltransferases or histone deacetylases. Unfortunately, these inhibitors are often non-selective... industry, the bioethanol industry, and the pharmaceutical industry, depend on the yeast, Saccharomyces..., including fermented beer and wine, fuels, pharmaceutical ingredients, and recombinant proteins. In order...