...Tethering dCas9 to the histone acetyltransferase p300 or the DNA methyltransferase DNMT3A allows researchers...2013). Enzymatic domains tethered to dCas9 often interact with DNA independently of dCas9 targeting (Galonska... independent of their enzymatic activity. They interact with transcriptional machinery to modify local...modifying other sites in the genome and directly interacting with transcription machinery.   Well, I’...demethylation?  Dividing cells depend on DNA methyltransferase 1 (DNMT1) to copy methylation marks from parental...   Let me leave you with a few practical considerations: Design your gRNA to overlap the CpG of interest...that unmethylated CpG so that nothing else can interact with it. Remember, the goal of this experiment...

...structural determinants of protein-DNA interactions and how these interactions regulation gene expression. Recent...structural determinants of protein-DNA interactions and how these interactions regulation gene expression.     ...are being awarded free plasmids form Addgene to accelerate their research. Read on to learn how these researchers.... These studies are expected to identify new therapeutic targets/solutions and enable scientists to discover...mutations. References: Baldwin, Enoch, et al. "Generation of ligand binding sites in T4 lysozyme by deficiency-creating...in brain development since 2000 and have shown several mechanisms by which they affect brain and muscle...muscle development. The Gronostajski lab has generated mice wherein it's possible to delete the functional...

...demonstrated that the caffeine concentrations in several beverages could be estimated by measuring growth of ...permits it to bind to the CIB1 (cryptochrome-interacting basic-helix-loop-helix) protein. After fusing...proteins of interest, a researcher can cause them to interact by stimulation with blue light.  Similarly, light...work including the soCoChR-GFP plasmid as well as several derivatives encoding alternative promoters and/...as starting material for the bioproduction of therapeutic methylxanthine drugs like theophylline. To facilitate... required enzymes, including a glutathione S-transferase homolog from a Janthinobacterium sp. Moreover...minimal media supplemented with various caffeinated beverages, but not with caffeine-free drinks. Finally, they...

...things) gene delivery, generating pluripotent stem cells, and developing gene therapies for monogenic diseases...excellent tools for genetic engineering. There are several different viral vectors to choose from, for example...which includes the gamma-retrovirus and lentivirus genera, have the unique ability to integrate permanently...separating viral genes and transgene expression across several plasmids. The most commonly used viral vector systems...of the lentiviral elements and the 2nd and 3rd Generation Lentiviral Systems can be found in our Lentiviral...gamma-retroviral packaging plasmids are not interchangeable. General envelope plasmids, such as VSV-G, may be used across...interest into this tetracycline inducible, 3rd generation lentiviral vector backbone from the Shih lab....

...needed for optogenetics. There are several important considerations with fabrication of the fiber optics...pulse generator such as the A.M.P.I Master-9 Pulse Stimulator, which can control output of several lasers...equipment, most of which is relatively cheap). Several protocols have been written outlining the different...stimulation. The scatter effect is also an important consideration when preparing the implant itself, which brings...least 85% of the initial laser output. The same considerations apply to the polishing and preparation of the... channel, Channelrhodopsin 2 (ChR2), exists in several states: closed/light receptive, open, closed/light...manipulation of the laser power source itself. Pulse generation (TTL pulse) is a standard electronics protocol...

...your serotypes, titer, and promoter. Note that several steps in testing are effected by multiple AAV variables... as proxies.  Finally, the size and receptor interactions of a serotype can affect spread from the injection...and are useful if you're going to be working in several different systems or want to transduce a large ...Various AAV Serotypes and Their Applications in Gene Therapy: An Overview. Cells, 12(5), 1–41. https://doi.org... Gene Delivery to the Hypothalamus. Human Gene Therapy, 21(6), 673–682. https://doi.org/10.1089/hum.2009.169...Carroll, S. J. (2019). Astrocyte-selective AAV gene therapy through the endogenous GFAP promoter results in... in the rat spinal cord following injury. Gene Therapy, 26(5), Article 5. https://doi.org/10.1038/s41434...

...cloning. The general cloning strategy described in the associated publication can be used to generate any combination... a modular mini-Tn5 vector that can be used to generate random mutagenesis libraries or to insert heterologous...oxidized. The Timer tool has already been used by several groups to monitor protein and cell ageing, as the...action via sequestration. The strategy relies on several observations: Rapamycin is a membrane-permeable...vectors are used with Multisite Gateway cloning to generate the final construct. Up to eight entry vectors...demonstrated the use of the SpyLigase peptide-peptide interaction to link affibodies or antibodies against common... Despite these advances, the Ting lab’s first-generation APEX suffered from inconsistent activity levels...

... plasmids for generating your own libraries are also available in the Sybody Generation Toolbox plasmid... two different coronaviruses into a llama over several days and isolated special camelid antibodies that...proteins that may unfold due to camelid high body temperature (37.2 - 39.2°C). The Seeger lab found a way around... around this by using plasmids to generate sybodies, synthetic nanobodies against the spike protein (Walter...blog post.  A rapid selection platform for generating sybodies against membrane proteins Developing...membrane protein of interest using the Sybody Generation Toolbox and commercially available reagents in...libraries (Zimmermann et al., 2018). Find the Sybody Generation Toolbox! Starting with libraries and a biotinylated...

...yeast and cell free expression systems. Overall the general strategy for protein expression consists ...or purify proteins to study their structure, interactions and functions. Proteins can also be purified...purified for use in molecular biology research (eg. polymerases and other enzymes might be purified and used ...derived from cells or using live cells. There are several types of expression systems used for protein production...Deciding which expression system to use depends on several factors: The protein you are trying to express...most popular hosts for protein expression with several strains that are specialized for protein expression...be purified from the cellular debris. There are several popular DNA vectors that can be used to produce...

...GFP/EGFP) for a transcriptional reporter or as a general cytoplasmic label of cells, there's no problem....to not produce artifacts in your experiments. Considerable effort has gone into making FPs monomeric, yet...ignorant of FP dimerization. Equally problematic, several reportedly monomeric FPs are not actually monomeric...more likely molecules of a particular type will interact. A protein with a nanomolar Kd will exist primarily... of the FP. The FP is confined to a plane and generally only rotates in 360°, increasing the probability...2]. 2. Fusion to an obligate dimer or oligomer Several cell proteins normally self-associate into homodimers... and increasing the signal from the second FP. Several early FRET biosensors were made with CFP and YFP...