We narrowed to 790 results for: Cre
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TypeBlog PostPublishedApril 20, 2023, 4 p.m....phobias–it’s a key part of being a scientist and an incredibly useful transferable skill. Being able to stand...even in my head. What came out, though, was an incredibly calm: “Excuse me, I seem to have lost my place...any silence, to talk. That moment taught me an incredibly valuable lesson: Your presentation will be better...your slide deck gets uploaded. Your mic emits screeching feedback. You’re muted on Zoom. Your video conferencing...provide quick updates during a company meeting increases visibility of your work, opens up collaboration...Conflict Resolution Transferable Skills Guide: Creativity...
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Making CRISPR Plasmids Using Fragmid
TypeBlog PostPublishedAug. 29, 2024, 1:15 p.m....2024). It reduces the amount of time to design and create a new vector to four days for an experienced bench...takes a lot of the design work out of the plasmid creation process. Each module of Fragmid represents a class...caption from McGee et al., 2024, used under the Creative Commons CC BY-NC-ND license. Once you have... system by sequencing 60 individual assemblies created over five months. When they tested those clones... plasmids were perfectly correct. Because they created the assemblies in duplicate, they were able to ...Doench lab share this new tool and hope it helps you create great plasmids for your next CRISPR experiment.... -
Viral Vectors 101: Chemogenetics
TypeBlog PostPublishedJuly 11, 2023, 1:15 p.m....further modified to create ultrapotent PSEMS (uPSEMs) that have drastically increased potency. Download ...signaling pathways and can lead to such responses as decreased heart rate (Redfern et al., 1999) or altered neuron...firing. Other DREADDs, such as the KORD receptor, increase this threshold and silence neurons. DREADD-activated...actuator modules, were developed with the goal of creating receptors with less endogenous activation and ... Higher potency means lower drug doses, which decreases the risk of off target effects. These synthetic... -
Overcoming the Challenges of Lentiviral Production
TypeBlog PostPublishedAug. 7, 2018, 12:31 p.m....consider scaling up by either increasing the number of 10 cm2 dishes or increasing the vessel size. Opting ...viral genome into additional plasmids thereby increasing the number of recombination events that are required...some cases, titers can drop 10-fold for every 2kb increase in insert size. To overcome this, lentivirus can... lentivirus routinely, you may opt for simply increasing the number of 10 cm2 dishes. While this option...handling multiple dishes will take longer and may increase the risk of mistakes or contamination. Finally... -
CRISPR 101: Multiplex Expression of gRNAs
TypeBlog PostPublishedSept. 10, 2020, 11:45 a.m....plasmid contains all of the desired gRNAs. This increases the chance that all the edits you want to make... cleave outside of their recognition sequence, creating flanking overhangs. These overhangs can be customized...Expression Systems (MuLE) Kit: This kit can be used to create lentiviral vectors expressing wt humanized Cas9...the gRNAs. gRNAs are flanked by glycine tRNAs to create polycistronic glycine tRNA-gRNA (PTG) constructs...using 3 distinct spacer-repeats, you could easily create 7 unique arrays (e.g. for spacers A, B, and C, ...is possible with efficiency >70%. CRMAGE is an incredibly fast protocol, with only 5 hours incubation time... Find plasmids from this paper at Addgene. 11. Cress, Brady F., et al. “CRISPathBrick: Modular Combinatorial... -
No Llamas Required - Synthetic Nanobodies Against Membrane Proteins
TypeBlog PostPublishedJune 18, 2020, 1:15 p.m....to, like ones that are hidden in the molecular crevices proteins on the cell surface. Figure 1: ...threonines at positions that can be randomized for creating the libraries (Zimmermann et al., 2018). Find ...protein (sybody), ribosome, and mRNA. During the screening process, displayed sybodies that bind to the biotinylated...the candidate sybodies in their coat protein to screen for binding to the target protein. The protocol...describes going through two rounds of phage display to increase the enrichment of target-specific sybodies. Final...The enriched sybody pools from the phage display screens are subcloned (using FX cloning) into the expression... -
Fluorescent Proteins 101: Fluorescent Protein Timers
TypeBlog PostPublishedMay 4, 2017, 2:30 p.m....Tsuboi et al.), limiting its use. Nonetheless, its creators noticed a few properties that have now become ... be. Cytotoxicity, improper localization, and decreased functionality are all possible consequences of...al.) Monitor gene expression in the developing pancreas by placing FP timer DsRed-E5 under the control...control of Neurog3 (a gene controlling pancreatic differentiation) in mouse embryos (Miyatsuka et al.) Distinguish...you planning on using an FP timer in a new and creative way? Let us know in the comments section below...differentiates human IPSCs into glucose-sensitive insulin-secreting beta-like cells." Nature communications 7 (2016... -
When is a Monomer not a Monomer? The Top Three Ways Your Favorite Fluorescent Protein Oligomerizes in Cells
TypeBlog PostPublishedApril 19, 2016, 2:30 p.m.... second FP, often decreasing the fluorescent signal from the first FP and increasing the signal from the...an FP to a transmembrane protein significantly increases the effective concentration of the FP. The FP ...to a plane and generally only rotates in 360°, increasing the probability that two FPs will collide with...polymer formation as fusion protein concentration increases. The polymerized fusion proteins that form as ... His interests include the quality control of secretory proteins in the Endoplasmic Reticulum and live... -
Summer SciComm Series: Modes of Communication
TypeBlog PostPublishedJuly 1, 2022, 1:15 p.m....well for everyone else? In truth, people are incredibly varied in how they prefer to consume information...matter will drive the kinds of communications you create. Wildlife biology and ecology can make for beautifully...Once you’ve answered the above questions, you can create your short list of options and then pick whichever...Medium to high Uses visuals, audios, and text. Incredibly immersive and flexibility to visualize and contextualize...format. Equipment, expense and expertise needed can increase exponentially with complexity. Podcasts Medium...audience and communicators. Well-developed content creation platforms. Algorithms can make it difficult ... -
pCXLE toolkit: Efficient episomal plasmid-based method to reprogram peripheral blood cells to iPSCs
TypeBlog PostPublishedDec. 14, 2017, 2:08 p.m.... I’ll describe an episomal vector system that increases reprogramming efficiency through the expression...requires stringent biosafety measures, thereby increasing the costs of the procedure. Episomal reprogramming...reprogramming is an ideal and cost-effective method for creating clinical-grade, safe, nonviral and nonintegrating...episomal reprogramming cocktail by Okita et al. increased the reprogramming efficiency in PBMCs [7]. The...transiently. The transiently expressed EBNA-1 increases protein expression from the other episomal plasmids...used on PBMCs, the pCXLE episomal vector system increases the efficiency of reprogramming from 0.001% to...