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  2. Expanding the Targeting Scope and Editing Efficiency of Adenine Base Editors

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    Blog Post
    ...in more contexts. References Gaudelli NM, Komor AC, Rees HA, Packer MS, Badran AH, Bryson DI, Liu DR...developed the Phage-Assisted Continuous Evolution (PACE) and Phage-Assisted Non-Continuous Evolution (PANCE...mutations, selection, and replications per day.  The PACE and PANCE systems use M13 bacteriophage, which rely...dilution step represents the major difference between PACE and PANCE: for PANCE the phage progeny is not diluted...-stringency PANCE, followed by higher stringency PACE, resulted in the evolution of TadA variants with...

  6. CRISPR Methods for Bacteria: Genome Engineering, CRISPRa, CRISPRi, Base Editing, and More

    Type
    Blog Post
    ...Streptomyces Species Using an Engineered CRISPR/Cas System. ACS Synth Biol 4:723–728 . https://doi.org/10.1021/sb500351f...for Modular Assembly of Broad-Host-Range Plasmids. ACS Synth Biol 7:1279–1290 . https://doi.org/10.1021/... most CRISPR-based genome engineering has taken place in other organisms. In many bacteria, unlike other...and chloramphenicol resistance pCRISPR: carries a spacer targeting the gene of interest and kanamycin resistance...template, and the locus cannot be recognized by the spacer-derived crRNA. However, if recombineering is unsuccessful...survive have inactivating mutations in the Cas9 or spacer sequence, and these rare events are easily detectable...successfully in Tatumella citrea, another species of Enterobacteriaceae, without the need for modification. This ...

  7. 22 Hot Plasmid Technologies from 2014

    Type
    Blog Post
    ...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...

  8. GPCRs: How Do They Work and How Do We Study Them?

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    Blog Post
    ...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 ...

  11. Viral Vectors 101: Chemogenetics

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    Blog Post
    ...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...

  12. CRISPR 101: Targeting Non-Coding RNAs with CRISPR/Cas9

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    Blog Post
    ...821. doi:10.1126/science.1225829  Hazan J, Bester AC. CRISPR-Based Approaches for the High-Throughput ...with multiple gRNAs poses the risk of disrupting adjacent genes, especially those regulated by or overlapping...al. 2019). The interactions between ncRNAs and adjacent genes are complex and not yet fully understood...

  13. A New Generation of Adenine Base Editors Improves Editing in Primary Human Cells

    Type
    Blog Post
    ...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...

  14. Truncated gRNAs for Regulating Gene Expression

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    Blog Post
    ...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...

  16. Getting to Know Expansion Microscopy

    Type
    Blog Post
    ...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 ...

  17. INTEGRATE: Bacterial Genome Engineering Using CRISPR-Transposons

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    Blog Post
    ...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...

  18. Better Dyeing Through Chemistry & Small Molecule Fluorophores

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    Blog Post
    ...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...

  19. CRISPR Between the Genes: How to Experiment with Enhancers and Epigenomics

    Type
    Blog Post
    ...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...

  20. Hot Plasmids: Fall 2024

    Type
    Blog Post
    ...Readout of Mechanical Forces in Biological Systems. ACS Sensors, 9(7), 3489–3495. https://doi.org/10.1021...
Showing: 41 - 60 of 721 results