...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...

...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....

...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...

...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...

...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...

...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...

... 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...

...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 ...

... 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...

...cytosine deaminase with Cas9 nickase (nCas9). nCas9 creates a nick in the DNA by cutting only a single strand...inhibitor (UGI) minimizes the second outcome and increases the generation of the desired T-A base pair. ...act on DNA for adenine base editing. They first created a defective chloramphenicol resistance gene (CamR...antibiotic resistance. To find such a protein, they created a mutant library of E. coli tRNA adenosine deaminases...into E. coli harboring the defective CamR gene. Screening for surviving colonies and subsequent rounds of...genotyping. If successful base editing causes the creation or destruction of any restriction enzyme (RE) ...

...humanities, and 32% in the social sciences go uncited. Creative titles and controversial topics can garner some...Article, since the 1980s researchers have been increasingly citing research from outside of their disciplines...interdisciplinary research while at the same time increasing interdisciplinary citations in their own work...instant sharing across multiple other sites), I created a new platform for sharing video abstracts about...effectively.” How to share your science through video Creating an effective video about your research is not ...offers a 5-step process and several examples for creating effective research videos.  Karen McKee, also...

...Saves Christmas! created by the Moore lab at the Van Andel Institute. Krampus was created by Nate Levine...Warrior The Moore lab, who was full of holiday creativity, also receives an honorable mention for their...2021   2nd Place: The Winter and the Falcon Tube created by the team efforts of the Yit Heng Chooi Lab at... December 4, 2021   3rd Place: O! Beaker Tree Created by Joanne Jary from Lab Support at the John Innes...

...DNA end of the break is resected by nucleases to create a ssDNA 3’ overhang. This will serve as both a ...considerations for designing a repair template to create mutations HDR donors can contain insertable sequences...circular plasmid templates is generally low; to increase the frequency of edits, researchers have designed...complementary ssDNA. Easi-CRISPR was shown to increasing editing efficiency from 1-10% with dsDNA to 25...Frequency of Homology-directed Repair (HDR) for Increased Efficiency of CRISPR/Cas-mediated Transgenesis...

...including easier automation, higher throughput screening, and a lower requirement for the amount of DNA...transgenic animals may choose Southern blotting to screen founder transgenic animals for the inserted transgene...DNA sequence content by comparing the fragments created by different restriction enzyme digests. Incubating...specific fragments of a DNA sequence of interest to create a physical map of the DNA. Starting in the 1970s...below), restriction mapping may still be used to screen multiple colonies before selecting a smaller subset...allowing for fluorometric-based detection. This increased sequencing speed ultimately leading to what are...

...donors to supply surplus lab equipment to carefully-screened research and teaching departments at universities.... Like Seeding Labs, Addgene also feels it’s incredibly important for scientific breakthroughs, reagents...scientific community and are always taking steps to increase plasmid distribution and disseminate knowledge... we need to expand our efforts to find new and creative ways to reach developing countries. As part of... eliminate much of white space on this map and increase plasmid sharing across the globe! We hope that...

... associated hybridoma cell lines. The DSHB was created by the National Institute for Child Health and ... which are each produced by a single B cell. To create a monoclonal antibody cell line, myeloma cells ...individually isolated and expanded. The clonal cells are screened to identify hybridoma lines, each producing a ...and protein tags (c-MYC and HA). We have also created the Monoclonal Antibody Research Institute dedicated...partially humanized rabbit antibodies that exhibit increased avidity to high profile antigens; and 4) generating...

...use the dropdown to increase the length of the list (purple oval in the screenshot below). You can also...plasmid page. Check out the purple arrow in the screenshot below to see what I mean. Additional features...list of papers citing your deposited plasmids Increased visibility of your plasmids in the "Associated...listed in the "Articles Citing This Plasmid" list increasing your work's visibility Help us improve our citations...

.... Figure 1: Diagram of AAV components. Created with BioRender.com. AAV vectors have both distinctive...AAV2, and so on. In addition, researchers have created many engineered serotypes by mixing, matching, ...been shown to transduce skeletal muscle, cardiac, pancreas, smooth muscle, CNS, kidney, and retinal cells...been shown to transduce liver, skeletal muscle, pancreatic, and retinal cells (Issa et al., 2023). AAV9 ...pUCmini-iCAP-AAV9.452sub.LUNG1 Goertsen et al., 2022 Pancreas AAV6, AAV8 Wang et al., 2006 Retina 7M8 Dalkara... J., & Xiao, X. (2006). Widespread and Stable Pancreatic Gene Transfer by Adeno-Associated Virus Vectors...

...specifies the target, the Cas9 endonuclease that creates the DNA double-strand break (DSB) at the target...knock in models). CRISPR mouse model basics To create a mouse model, the gRNA, Cas9, and donor oligonucleotide...selection offered by mouse ESCs. For example, we often screen only 15-25 mice when generating knockout models...for generating mouse models Although CRISPR is incredibly useful for generating mutations by NHEJ and generating...CRISPR, one must beware that not all gRNAs are created equal. Some work better than others. There are ...Mice with Extensive Gene Humanization (25-kbp) Created Using Embryonic Stem Cell/Blastocyst and CRISPR...

...information on black screens, but not to know even the life cycle of the creature they deal with.” So ...Wildlife Magazine. It is easy to see how these incredible images are attracting so much attention. When...efforts, as the subject matter is seen to have increased value and is appreciated by a wider audience for... by holding scientific art competitions where creations can go on display for all to enjoy. The largest...