...classification. Lentivirus (a type of retrovirus) and gamma retrovirus require more caution as they are integrating...virus and its infectivity in check. Engineering controls, safety systems that are built into how something...functions, were designed for this purpose. One such control is dividing up the essential components of a virus...virus into different plasmids. For a retroviral system this looks like the genetic cargo in one plasmid... researcher.   Figure 1: Separation of retroviral packaging components onto distinct plasmids for...common types of viruses grown in the lab are gamma retrovirus, lentivirus, adenovirus, and adeno-associated...  Susanna Stroik...

... No spatiotemporal control. Many PPI detection methods lack spatiotemporal control or an “on/off switch...identifying PPIs, such as affinity purification mass spectrometry (AP-MS), lack the ability to specifically identify... by creating Split-BioID, a spatiotemporally controllable version of the proximity-dependent biotinylation...method Enter Split-BioID, a spatiotemporally controllable version of the BioID protocol. The critical ...interaction partners if they are enriched relative to controls using Split-BioID-GFP and BioID run on six unrelated...combinations and pick the conditions that give the strongest biotinylation signal, as measured via Western ...methods, including BioID, is its spatiotemporal controlled labeling of interacting proteins; nearby proteins...

...delivery include plasmid transfection, RNP electroporation, and viral transduction; but these methods ...gRNAs. Viral envelope proteins To alter the cell tropism of Nanoblades, different viral envelope protein...with a mix of VSV-G and the baboon endogenous retrovirus Rless glycoprotein (BaEVRless) had high rates...gene, while editing was not detected in cells electroporated with RNPs using the same guide. Nanoblades ...influence the level of off-target effects, with electroporation of RNPs generally having lower off-target edits...polymer. Polybrene has long been used to increase retroviral transduction efficiency by neutralizing the charge...the cell surface, and also helps form DNA and retroviral-particle or VLPs complexes. The team found that...

... Overview of in vitro AAV CRISPR screens. Created with BioRender.com. The in vitro option requires... screens may be AAVantageous!       In vivo, in vitro, ex vivo CRISPR screens can be carried out using...using a few different methods — in vivo, in vitro, and ex vivo. The general process of these screens is the...with transducing cells with the CRISPR library in vitro before transplanting them into mice. For in vivo...when designing screens. Each serotype has its own tropism, or a preference to infect specific cell or tissue...tissue types. Using serotypes that have a known tropism for your cells of interest — for example, AAV8 ... used these psuedotypes to further improve the tropism of AAV vectors, enhancing their affinity for specific...

... APC control, with APC and APC-CY7 plotted against each other. Since it is a single-color control, you...example with single color control. Flow plots showing an APC single-color control sample before (A) and after...recommended to start compensating with single-color controls before moving onto your experimental samples. ...samples without needing to use your single-color controls as a starting point. Applying compensation to ...experimental samples as they did with your single control samples: if the population needs to be shifted ...School.   More resources on the Addgene blog Introduction to Gating in Flow Cytometry Antibodies 101: ...

...action potentials, muscle contractions, and the control of voltage gated ion channels all depend on transmitting...been successful even at the single neuron level in vitro. In vivo monitoring has also been achieved with ...effective at the single cell level as they are in vitro. Factors like photobleaching, lack of red-shifted...rhodopsin. FP-fusion to rhodopsin resulted in electrochromic FRET capable GEVIs, with rhodopsin quenching...membrane as well. Another flavor of these tools is Voltron, which uses rhodopsin as the voltage sensing component...genetically encoded component is delivered in vivo or in vitro and a small molecule is added exogenously to measure...Optogenetics Fluorescent Proteins 101: Introduction to FRET ...

...lab-adapted bacteria because it has been challenging to introduce CRISPRi machinery into diverse bacteria species... gRNAs, and dCas9 proteins. Second, it can be introduced into a broader range of bacteria. Once transferred...., 2019 with permission.   Two methods to introduce Mobile-CRISPRi by bacterial conjugation For gram...gram negative bacteria, Mobile-CRISPRi is introduced using the bacterial Tn7 transposon system. This system...firmicute gram positive bacteria, Mobile-CRISPRi is introduced using the bacterial integrative and conjugative...Mobile-CRISPRi. The Mobile-CRISPRi ICE element is first introduced into the B. subtilis genome. Mobile-CRISPRi is...into the trnS-leu2 locus.   Figure 2: Introducing Mobile-CRISPRi into bacteria. Top: A Tn7 transposon...

...AAV PHP.eB Controls Wilson New viral vector pENN.AAV.EF1a.eGFP.WPRE.rBG AAV PHP.eB Controls Wilson New...pAAV-CaMKIIa-EGFP AAV PHP.eB Controls Roth New serotype pAAV-hDlx-Flex-dTomato-Fishell_7 AAV2 Controls Fishell New serotype...  AAV PHP.eB Controls Gradinaru New serotype pAAV-CaMKIIa-mCherry  AAV PHP.eB Controls Deisseroth New...New serotype pAAV-Ef1a-mCherry  AAV PHP.eB Controls Deisseroth New serotype Looking for more viral...

...unknown sample. To do this, you serially dilute a control sample of known concentration and measure a specific...example, a BCA assay uses a serial dilution of a control protein, incubated alongside an unknown sample ... absorbance of both is then measured on a spectrophotometer and concentration versus absorbance is plotted... a standard curve by serially diluting a known control sample, called a standard. The ideal standard curve...curve accordingly.    Figure 1: A 1 mg/mL control is diluted in a series ranging from 1:2 to 1:32... for each run. To validate, include a positive control of known concentration in the experiment and use...  Include both a standard curve and a positive control in every run.  When setting up your dilution series...

...only a few, very strong associations.   Unexpected results Their results showed 96 strongly correlated genetic...may be the result of a technical artifact that introduced an ancestry bias into their experiments instead... prevent Cas9 from cutting, both of which can introduce significant bias into the results. But the reference...mapped to a SNP would be ignored. Working with Lia Petronio and Katherine Huang from Broad’s Pattern team,... of ancestral bias into genome-wide screens, introducing bias into all work based on these and any other...an ancestry group what is actually an artifact introduced by bias within the design of the experiment or...Noorbakhsh, Isabella Boyle, Joshua Dempster, Lia Petronio, Katherine Huang, Alham Saadat, Thomas Green, ...