...transfer plasmid with the E1 and E3 (Early 1 and Early 3) genes deleted. These genes’ functions are rescued...Degeneration in Tissue Culture. Proc Soc Exp Biol Med. 84(3), 570–573. https://doi.org/10.3181/00379727-84-20714...this introduction gives a sense of what AdVs are about – and we hope that it inspires further use of existing...

...Originally published Mar 3, 2016 and last updated Apr 13, 2021 by Will Arnold. Although CRISPR systems...a subsequent curing protocol that requires only 2-3 hours incubation. E. coli and T. citrea scarless editing...the QCascade DNA-targeting module with the crRNA, (3) three transposase proteins, and (4) the donor DNA...CRISPR Featured Topic Page Learn more about Cpf1 Find out about CRISPR Multiplexing Resources on Addgene.org...coli and other bacterial species. Read on to learn about the tools available for bacteria and some of the...that can be used in this way. As described in the above CRISPRi section, the Marraffini Lab published a ...as a collection of organisms, their genes, and metabolic processes, occupying the gut. Many human diseases...

...228, no. 2 (2012): 267-75. PubMed PMID: 22767332.  3. Kim, Dohoon, Chun-Hyung Kim, Jung-Il Moon, Young-...PubMed PMID: 19337237. PubMed Central PMCID: 2677165. 3. Jia, Fangjun, Kitchener D. Wilson, Ning Sun, Deepak... deriving human iPS cells." Nature Methods 7, no. 3 (2010): 197-99. PubMed PMID: 20139967. PubMed Central...methods do you have experience using? Let us know about your iPSCs experiences in the comments below! References...Additional Resources on the Addgene Blog Learn about Methods to Reprogram Peripheral Blood Cells Check...

...on the future of the scientific endeavor October 2-3, 2014 Boston University Jacob Sleeper Auditorium, ...doing science in some form? There is a lot of talk about the state of scientist training in the US and around...

...individual, sharp and evenly spaced peaks …like these…. 3. Expect to get 500-700 bases of clean reliable DNA...and troubleshooting your DNA sequencing results Above (top right corner of this post) is an example of...

...are 24 variants of the cas14 gene that cluster into 3 subgroups (cas14a-c). All these variants share the...Additional resources on the Addgene Blog Learn more about DETECTR Browse all of the CRISPR blog posts Download...

...Method with High Throughput Capability." PLoS ONE. 3(11): e3647 (5 November 2008).  ...protein,” he says. “There is nothing you can do about it but be gentle. If it no longer works, just trash...

... MD but later got obsessed with research. Chapter 3: How all basic research ends up being translational...promote scientific sharing. Listen to learn more about Darrell, his research, and his focus on open science...Medicine and in the Department of Pathology and Laboratory Medicine. He attends in the Medical Intensive...NIH-funded Principle Investigator in the Kotton Laboratory and is the founding Director of the Center for...Podcast.     Additional Resources from Addgene Learn about Retinal Gene Therapy from Connie Cepko Find Plasmids...Plasmids from the Kotton Lab Learn more about Stem Cells ...

...with an isotype-matched control.      Figure 3: A preimmune serum control addresses situations where....” British Journal of Pharmacology, vol. 175, no. 3, Wiley, 19 Jan. 2018, pp. 407–411. Crossref, doi:10.1111...subjected to the experimental condition. In the example above, a negative control could be a third group of mice...in the experimental protocol. In the experiment above, you could repeat the tau western blot several times...Baseline secondary signaling levels In addition to the above-mentioned safeguards, you’ll need to test the secondary...Flow cytometry In addition to the controls listed above, viability and compensation controls are recommended...technique before starting. For more information about antibodies and antibody-based experiments, check...

...working with RNA and extracting RNA without a kit. 3. SARS-CoV-2/COVID-19 Detection Methods Based on CRISPR... highlighted reagent repositories and other collaborative efforts in the scientific community to speed...store my plasmid in TE, Tris, or water? And what about temperature? Find the details in this blog post....dominates DNA quantification discussions, but what about the other methods to quantify DNA? Here we cover...Incompatibility by Leah Schiesow If you're thinking about hosting multiple plasmids in one cell, it might ...

...available as viral vectors (here and here).  Figure 3: JEDI-2P was identified from a high-throughput screen...microscopy. Cell. 2022 Aug 16. pii: S0092-8674(22)00916-3. doi: 10.1016/j.cell.2022.07.013 pYtags illuminate...screen (Koveal et al. 2022). Lactate is a key metabolite in glycolysis, so this sensor will be a valuable...valuable tool to study cellular physiology and metabolism under various conditions. LiLac has improved response...going viral! In addition to the viral preps noted above for iGluSnFR3 and JEDI-2P, several biosensors already...innovation next year! Did we miss any? Let us know about your favorite new biosensor plasmids in the comments...

...perform downstream characterization of, isolating 2 or 3 distinct clones to perform key experiments on is a...CRISPR-Cas System in Gene Editing. Mol Ther. 2021 Mar 3;29(3):937-948. doi: 10.1016/j.ymthe.2020.11.009  Guo C,...is an off-target effect. Should I be concerned about off-target effects? Depending on your experimental...this could be a huge concern, or nothing to worry about all. Let’s consider several scenarios below for ...Predicting off-target events As mentioned in the above section, there are gRNA softwares available which...diligence to ensure they are indeed a non-issue. Above we mentioned how to predict off-target frequency...

...Kinases: they regulate many proteins, with ~1/3 of human proteins predicted to be phosphorylated on at...Blog Learn about photoswitchable kinases Learn more about fluorescent biosensors Learn about using phosphoserine...

... guest Drew Endy, in celebration of open science. 3)      Also in June, the G8 Science Ministers issued...study probably doesn’t say as much about open access as it does about the state of peer review. 8)     ...University Open Data Access (YODA) Project, “a collaborative endeavor guided by aspirations to promote open...

... it from carrying out its proper function (Figure 3).  The Hehnly Lab used such a strategy to dissect ...injection of plasmids and other materials.    Figure 3: Optogenetic inhibition of Rab protein vesicle trafficking... Check out our Optogenetics Guide to learn more about how these modules work, and read on to learn how...to the cell membrane isn’t powerful enough, what about altering the chemical composition of the membrane...

...to allow for antibodies to enter the cell (Figure 3). A useful side effect of the fixing/permeabilization...cytosolic procedure in that case.     Figure 3: For intracellular labeling cells need to be fixed..., you may want to look at other methods. In the above cases, a good alternative is using a two-protein... region of the primary antibody (Figure 1b). As above, the advantage of this system is increased flexibility...

... et al. designed gRNAs to target the HIV-1 5’ and 3’ LTRs and expressed them along with Cas9 in the T-...eradicating HIV-1/AIDS. J Neurovirol. 2015 Jun;21(3):310-21. PubMed PMID: 25716921. Additional Resources...reservoirs, chronic inflammation, and negative metabolic effects of ART. Why can’t the body mount an immune...

...Blog (other programs are discussed below). Chpater 3: SITN Popularity and Events - Amy discusses some of...To learn more about the STIN's podcast, email [email protected]    For more info about starting your...evens where scientists can grab a beer and chat about their work at a local bar. It doesn't have to be...1: Introduction - Vini and Amy tell us a little about themselves. Chapter 2: What is Science in the News...science communicators. In this chapter we learn about some of their events - science seminars, science...the lab and working with SITN. She points to collaboration and delegating as keys achieving success. Chapter...your own SciComm organization and to learn about some of SITN's other initiatives, email [email protected]...

...for making scientific graphics. PS: they’re free. 3. Fluorescent Proteins 101: GFP Fusion Proteins - Making...mNeonGreen, mTFP1, and mWasabi by Jennifer Tsang Learn about mNeonGreen, mTFP1, and mWasabi and find plasmids...Broccoli, Spinach, and Corn, for example. Learn more about the aptamer soup in this blog post. 5. Five Popular...

...similarly but not identically to BE3, modifying a 3-5 base window 18 bases upstream of the PAM.  David...than BE2 Nishida et al., 2016 Target-AID   Edits 3-5 base window surrounding -18 position upstream of... efficiency and product purity. Science Advances, 3(8), eaao4774. https://doi.org/10.1126/sciadv.aao4774...Homology-Directed Repair Learn about base editing for plants Read about four base editor reporters Additional... The BE3 system increased editing frequency to above 30% for a variety of targets in human cell lines... Liu lab created BE4max and AncBE4max mentioned above, they also made an adenine base editor with improved...cell therapy applications. Dual base editors What about combining the function of both editors into one?...