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We narrowed to 1,006 results for: RING

Showing: 241 - 280 of 1006 results

  1. Stabilized Bacterial Promoters: Constant Gene Expression at any Copy Number

    Type
    Blog Post
    ...for many metabolic engineering and synthetic biology goals. In metabolic engineering, the expression of...interfere with the precision needed in metabolic engineering or other synthetic biology circuits. What is ... a look at how the stabilized promoters perform during these types of fluctuations. When cells are dividing... can also cause changes in plasmid copy number. During the shift from exponential to stationary phase,...promoters were able to better modulate gene expression during shifts in growth rate as compared to a constitutive...To simulate the increase in plasmid copy number during the transition to stationary phase, Segall-Shapiro...they could alter copy number of the plasmid by altering IPTG concentrations. Higher IPTG concentration...

  2. Delivery Methods for Generating iPSCs

    Type
    Blog Post
    ...leads to incomplete reprogramming. Additionally, lingering expression or re-expression of viral transgenes...temperature sensitive Sendai virus which is removed by culturing cells at 38℃, but screening cells for the presence...considered the safest since there’s minimal risk of lingering expression of reprogramming factors. Non-replicating...the plasmid in mammalian cells and assists in tethering the vector to the cell’s chromosomes. With the... high marks for safety.   Protein Delivery: Delivering reprogramming factors as protein is another way... week for 6 weeks, and still required further culturing to generate iPSC colonies. Reprogramming efficiencies...three germ layers (ectoderm, mesoderm, endoderm). Measuring pluripotency of mouse cells typically involves...

  3. CRISPR/Cas9 FAQs Answered!

    Type
    Blog Post
    ...makes sense that scientists, many of whom are venturing into genome editing for the first time, have lots... the growing PubMed article list, the Genome Engineering Using CRISPR/Cas Systems has expanded out of ...wildtype or double nickase for my CRISPR genome engineering experiments? A1: When assessing which nickase...nickase type to use for your CRISPR genome engineering experiments, consider that wildtype Cas9 with optimized...amazing resources for the scientific community and sharing these resources with Addgene. Thank you, Le! This...K., & Carroll, D. (2013). Donor DNA Utilization During Gene Targeting with Zinc-Finger Nucleases. G3 Genes... L. A., & Zhang, F. (2013). Multiplex genome engineering using CRISPR/CAS systems. Science, 339(6121),...

  4. An “elegans” Approach to Better CRISPR/Cas9 Editing Efficiency

    Type
    Blog Post
    ... organisms which I use to study gene regulation during developmental processes, CRISPR/Cas9 allows us ...laborious handling (screening 768 F1 animals). Recovering rare editing events in a sea of unedited animals..., which makes screening extremely rapid. This stringent selection allowed me to optimize a range of editing...al., 2018). They further built on this work, discovering that melting and rapidly cooling dsDNA repair...also found that the majority of knock-ins were occurring 24 hours post-injection, making screening for ...where the 5’ ends of the PAMs are facing outwards, ensuring the PAMs remain on the chromosome after DSB formation...Toolkit and SapTrap-SEC kit for C. elegans genome engineering have also facilitated CRISPR editing in C. elegans...

  5. Truncated gRNAs for Regulating Gene Expression

    Type
    Blog Post
    ...the Wyss Institute for Biologically Inspired Engineering. CRISPR/Cas9 technology has revolutionized the...the fields of molecular biology and bioengineering, as it has facilitated the development of a simple ...relative nuclease efficiencies as well as to more stringent targeting rules that result in a decrease in available... as a means of modulating gene expression (6). Altering the length of gRNAs targeting different sites ...Santa Cruz, where her research is focused on engineering nanoparticles as drug delivery systems. She is...develop several popular CRISPR-based tools for altering DNA sequence or regulating gene expression.   ..., Raj, et al. "Unraveling CRISPR-Cas9 genome engineering parameters via a library-on-library approach....

  6. DNA Purification Without a Kit

    Type
    Blog Post
    ...proteins in solution. The pH is then lowered using a renaturing solution, which causes the proteins and genomic...purification and DNA gel extraction with glass syringe filters Starting material: bacterial cultures or...binding DNA to silica columns, it’s bound to glass syringe filters. Normally DNA does not bind silica or glass... kit takes 1-1.5 hours to complete, while this syringe-based method can be completed in 20 to 30 minutes...culture you’re working with. Overall, the glass syringe filter method is not only cheaper, but also faster...gel slices Product: DNA fragments You might be wondering, “How is milk going to help me extract DNA from... 48 kb with high yields and without degradation during the extraction procedure. Additionally, purified...

  7. CRISPR 101: Ribonucleoprotein (RNP) Delivery

    Type
    Blog Post
    ...make genomic alterations, bringing about a revolution in genome engineering, with new techniques rapidly...and can be used for most of the current genome engineering applications of CRISPR including: generating ...delivery. You’ve decided to use RNPs, now what? Preparing the Cas9-gRNA ribonucleoprotein The first step... the Jacob Corn and Alex Schier laboratories. Delivering Cas9-gRNA ribonucleoproteins  An advantage of...applications. This technique utilizes Cas9 proteins harboring receptor ligands (C in the figure above), which...stable in human plasma than the standard spCas9 (Harrington et al. 2017). Lastly, as with any scientific ... Addgene.  Andrew's interests include genome engineering and microbiology.     References 1. Chen, Sean...

  8. Reflections and Looking Ahead

    Type
    Blog Post
    ...accelerating scientific discovery. Over 150,000 plasmids empowering researchers worldwide. Countless breakthroughs... to make it easier to find what you need and to bring it in line with current Web Content Accessibility...contributions to key research papers are already inspiring higher standards in antibody use (Ayoubi et al...packaged on demand in five distinct serotypes, offering you unparalleled flexibility. But this is just...characterization data for over 150,000 plasmids, empowering bioinformaticians, data scientists, and developers...collaboration formed, and every breakthrough enabled brings us closer to making an impact on humanity's greatest...this incredible journey. Together, we're not just sharing research materials — we're accelerating the future...

  9. Management for Scientists: Seeking Feedback

    Type
    Blog Post
    ... Members periodically break, go exploring outside the team, and bring information back. No team has all...contribution short and sweet. One key to this is ensuring that the group always conducts exceptionally well-managed...manager can be efficient without delegating and empowering the team to solve problems together (tactics ...important paths of information flow is the one that brings feedback back to the manager. Team members rarely...often A one-time event doesn't work to get people sharing.   Frequently ask direct questions for specific...at formal communications events or at informal gatherings, like at lunch. Many experts recommend using ...constructive feedback and suggestions. Consider offering rewards (gift card, afternoon off, etc.) Act on...

  10. Transferable Skills Guide: External Collaborations

    Type
    Blog Post
    ...transferable skills while working in the lab and by offering advice on areas where you might need some help...important in a non-research context, especially when considering small to mid-sized companies and organizations...time I’ve had a role in organizing collaborations. During my graduate studies at Karolinska Institutet I ...what was expected, and keeping track of speakers. During your graduate or postdoctoral work there are loads...you develop will lead! Read our blog post about fostering communication in remote collaborations! Stay in...Great progress! You are collaborating with your neighboring lab, you've met interesting people at conferences...twitter and maintain a connection by commenting and sharing. LinkedIn or ResearchGate allow you to send personal...

  11. Healthcare Consulting: A Door to the Business of Life Sciences

    Type
    Blog Post
    ... your foot in the door When I was first considering preparing for and transitioning to a career in healthcare... Healthcare consulting is a fast-paced field, requiring people who are willing to quickly learn, and apply...governments on the most effective and efficient ways of delivering healthcare. Clients typically engage consultants...fields. I got my PhD a couple of years ago in Bioengineering, and one of the things that pleasantly surprised... suite of benefits a given therapeutic approach brings to patients, doctors, and insurance providers, ...aspect of research in biology or medicine you will bring an essential set of tools to a consulting team. ...an increasing recognition of the value graduates bring to these roles. Healthcare consulting is a demanding...

  12. 6 Steps to Submitting a Resume That Gets Seen

    Type
    Blog Post
    ...Read here to get help if you are having trouble preparing your most excellent resume. 2) Follow job boards...reviewed regardless of personal connections to the hiring group.  Read here for more on timely submission... job description is usually a wish list and the hiring manager knows that no applicant is likely to meet...Your resume must be in the official “system” for hiring managers to take next steps and follow up especially... as you will not likely know the exact person monitoring submissions (usually a Human Resources representative... your resume pdf document. 6) Use your network! During the entire process, use whatever means you can ...can to get your resume into the hands of the hiring manager on paper or as an email send directly from a...

  13. An Addgene Summer Internship

    Type
    Blog Post
    ...C., Addgene’s intern coordinator shortly after hearing her talk about Addgene’s internship opportunity...summer interns presenting their capstone projects during the Summer Intern Poster Presentation.  As ...the Syntaxin3 gene in a pLenti vector backbone. During those five weeks, I worked with my mentors, Katherine...always came with surprises. Originally we thought during the first three weeks that we had made a mistake...not having the right concentration of a reagent during our HiFi assembly but decided to go ahead anyway...restart the procedure we would not have enough time. During the following two weeks, we were able to obtain...on my capstone project report and presentation, during the final week at Addgene. This report shared my...

  14. Technical Design of a Western Blot

    Type
    Blog Post
    ... assays are not with denaturing agents like DTT or β-mercaptoethanol.  Denaturing proteins To denature...blot, your loading buffer will not contain any denaturing agents. However, you’ll still have to lyse your... post, we are assuming that you are running a denaturing blot.  There are a number of lysing methods and... aliquot your sample(s) before adding in your denaturing and/or loading buffers, as most Bradford assays... proteins in different patterns, due to their differing chemistries. You therefore cannot compare Bis ...and reprobed multiple times. It doesn’t degrade during long-term storage and has both high binding capacity...well for proteins of most sizes and are worth considering if your proteins of interest vary wildly in size...

  15. Viral Vectors 101: Integrase-Deficient Lentiviral Vectors

    Type
    Blog Post
    ...research purposes, like creating stable cell lines, delivering CRISPR/Cas9 to cells for genome editing, and ... host’s genome isn’t uncommon. Many naturally occurring viruses occasionally integrate their genomes into...the host genome via the viral proteins packaged during production (read our blog post on the retroviral...time, IDLVs are ideal. As the episomes are lost during cell division, the expression of the delivered ...production protocol.  One thing to keep in mind is that titering these IDLVs may require a slightly different protocol...regular lentiviral vectors. This is because most titering methods rely on isolating genomic DNA from transduced...DNA along with a reference cellular gene. When titering IDLVs, there is no integrated piece to measure...

  16. Cultivating Community Science at BosLab

    Type
    Blog Post
    ...be taken seriously so we made it official by registering BosLab inc. as a nonprofit organization in the...are skilled with audiovisual media, software engineering, business consulting, mathematics, and more. ... to perform proof-of-concept experiments for discovering novel therapeutics. Dave is a chemical biologist...and in his free time, he is pursuing a plant engineering project at BosLab. Once he optimizes plant cell...knowledge with the community by teaching a DIY plant engineering workshop. Community projects at BosLab Not all...and Mrina Nikrad have started the Open Yeast Engineering Project (OYEP). Through this project, community...do similar things. The focus is to make yeast engineering accessible to everyone but these projects could...

  17. Finding Your Perfect Job After University

    Type
    Blog Post
    ...problem solving. I believe that is something lacking during degree courses; they never really prepare you for...failure and problem solving scientists experience during research. Working abroad in rural Africa Once this...another area which I was really passionate about during my degree. I loved parasitology. However, when ... held me back; I do not have a second language. During the recession there were many qualified scientists...Genetics to give me an edge over other graduates during the recession. As each advertised job received ...the training and experience relating to good manufacturing practice (GMP) and good lab practice (GLP) (...recognised the logo, but didn’t know much else. During the interviews, I asked if I could contribute more...

  18. It’s Bioelectric! An Exciting Interdisciplinary Field of Research

    Type
    Blog Post
    ...head when you hear the phrase “action potential?” Firing neurons are likely the first thing that comes to...reprogrammed regenerating tissues without using genome engineering. Planarian worms cut into three segments “remember...communication is briefly inhibited in the entire worm during regeneration, the worm creates two heads. Subsequent...resources from the bioelectricity field In addition to sharing tools to study bioelectricity, bioelectric researchers...defects, regeneration, medicine, and synthetic bioengineering. References Chernet, Brook T., et al. "Use...growth and form." Annual review of biomedical engineering 19 (2017): 353-387. PubMed PMID: 28633567. Mathews...bioelectricity for regenerative and synthetic bioengineering." Current opinion in biotechnology 52 (2018...

  19. Addgene: The First Twenty Years

    Type
    Blog Post
    ...open science nonprofit that would facilitate the sharing of plasmids between scientists. The service would...resource. I really appreciate the work you do to make sharing reagents and protocols so easy!” — Nina Jain, Harvard...seven before we had 1,000 depositing scientists. Ensuring that people knew how to access Addgene, and that...2012, our repository was rapidly able to support sharing of CRISPR constructs and information across the...lentiviral and AAV vectors, as well as developing and sharing supportive educational resources. By 2017, we had...NGS run.   To twenty years and beyond! Which brings us to 2024, Addgene’s 20th anniversary and the ...been made because scientists believe in openly sharing their plasmids. It’s been a privilege to find and...

  20. Gendered Innovations: Why Does Sex of the Cell Matter?

    Type
    Blog Post
    ...numbers of women participating in science and engineering. “Fix the Institutions” promotes gender equality...billions—but when drugs fail, they cause human suffering and death. Doing research right has the potential...between male and female cells. The result of not considering the sex of the stem cells was that their male...sex, and it’s important to analyze these factors during an experiment. In the case of stem cells, these...tissues—when that information is reliable—is important to ensuring excellence in basic research.  In addition to ... the sex of the most commonly used cell lines appearing in American Journal of Physiology-Cell Physiology...clue or potential place to start research is by considering sex disparities in disease severity. Such disparities...

  21. Antibodies 101: Stripping and Reprobing Western Blots

    Type
    Blog Post
    ...wash buffer such as PBS or TBST. Stringent stripping The stringent stripping solution uses heat and detergent...your sample? Or what if something weird happened during your western and your results look… funky? Do you...along with the antibodies, especially with more stringent stripping methods. If you are planning to detect...that section of the blot unusable for reprobing. Preparing your western blot Run your gel and membrane transfer...primary and secondary antibodies from your membrane, clearing the way for the second blot. There are multiple...with a mild stripping solution and move onto a stringent solution if the mild is not effective. You may...perhaps using longer incubation times, or try the stringent solution if you started with the mild one. Reprobe...

  22. Multiplexed Capture of Promoter-enhancer 3D Chromatin Structures Using CRISPR

    Type
    Blog Post
    ... steps, until its connection with DNA is broken during cross-link reversal (not shown in diagram). Cross-links...sequence targeted in the experiment. Figure 2: Comparing CAPTURE and CAPTURE 2.0.     Original CAPTURE...) one or more gRNAs Two separate lentiviruses delivering 1) a BioTAP-tagged dCas9 and an eGFP tag, and...detecting chromatin interactions opens the door for answering more questions about chromatin conformations. ...‘social networks’ of 157 super enhancers.  3. Measuring temporal changes in chromatin interactions A third...in chromatin conformation, like those that occur during development and cellular differentiation. Using...the Xu Lab tracked enhancer-promoter interactions during erythroid differentiation. CAPTURE 2.0 captures...

  23. AAV Vector Quality Control: Going the Extra Mile with NGS

    Type
    Blog Post
    ...packaged inside the vectors during production. These impurities cannot be removed during the purification step...recombinase-independent recombination during plasmid growth in bacteria or during viral production. This recombination... control assays for AAVs. These steps include measuring viral titer and assessing purity. But Addgene ...wrong plasmid? What if bacteria snuck into the prep during the production process? These questions are especially...materials. Addgene is going the extra mile and pioneering the systematic use of this new QC assay to guarantee...Evidence for Encapsidation of Prokaryotic Sequences during Recombinant Adeno-Associated Virus Production and...

  24. Tips for a 1st time AAV user (by a Rookie AAV user)

    Type
    Blog Post
    ...of choice is AAV, with nearly every experiment requiring AAV. Before joining my lab, I had never worked... a heated stir plate to dissolve and requires monitoring so that the solution doesn’t get too hot because.... Rather than investing all of this effort in preparing reagents, I choose to only harvest the cell pellet...layered and then the viral suspension generated during AAV production is overlaid. Following ultracentrifugation...first purification. This helped me get a feel for layering the iodixanol solutions and let me double-check...make one or two small aliquots to use for qPCR titering. This helps avoid multiple freeze/thaws of your...of virus. AAV titration by qPCR Overview: AAV titering by qPCR quantifies the number of genome-containing...

  25. Experimenting with New Careers while in Grad School

    Type
    Blog Post
    ...path began to take shape, was when I started volunteering. I wasn’t sure yet what a career in science ...volunteered at a place that made me happy. I started volunteering in the “Dinosphere” almost every week. I gained...dinosaurs, and met wonderful people. When I started volunteering, I was worried about becoming one of those angry...t sure what I wanted to do. A few weeks into volunteering, I was talking to a little girl about fossils...Beyond the outreach experience I gained from volunteering, it gave me a place to go once a week to get...before founding CINSO. Now, the network I built during graduate school is my informal advisory board. ...to share valuable life lessons they have learned during their scientific training and wrote about them ...

  26. Which Fluorescence Microscopy Techniques is Best for Me?

    Type
    Blog Post
    ...the palm of your hand. Although white light is entering your hand, only orange/red light can be seen exiting...optical section is achieved that boosts contrast by filtering out emission light from fluorophores higher in...as spinning disk confocal. However, due to the scattering nature of biological tissue, imaging more than...parallelization (7) have allowed for real-time monitoring of neuronal activity across substantial 3D volumes... allow for multi-view imaging can image thick scattering samples in their entirety, something that is ...excitation light which can lead to photobleaching when acquiring multiple images over a large axial range. 2-photon...overcome this (8). The recent development of tissue clearing techniques (9) now allows researchers to image...

  27. Tips for arabidopsis transformation

    Type
    Blog Post
    ...they begin flowering. It is important to perform the transformation not long after flowering begins. If...RFP in the seed (Emami, Yee and Dinneny, 2013). Preparing plants for transformation Whatever plants you ...you can cut off the primary inflorescence (the flowering portion of the plant) near the base of the stem...al., 2006. Tips: Don’t plate seeds too densely during selection: this will increase background growth... (2002) Arabidopsis : a laboratory manual. Cold Spring Harbor Laboratory Press. Available at: https://...). Additional resources on the Addgene blog Engineering the plant genome using CRISPR/Cas9 Using pSiM24...pSiM24 for simplifying plant genetic engineering Using arabidopsis in the classroom Resources on Addgene.org...

  28. How to Deposit Your Plasmids with Addgene

    Type
    Blog Post
    ...clicking “Add Plasmid to this Article” (left) and entering a plasmid name,plasmid type, and plasmid description...the plasmid names and descriptions first before entering the data. This step associates an Addgene ID with...plasmids all built with the same backbone and only differing in the gene/insert. Pooled libraries are normally... should click the "Enter Data" button to begin entering the specifics of your plasmids. Figure 5: Click... plasmid information. Enter plasmid information During the remainder of the process, you will enter plasmid.... You can complete these pages sequentially by entering the appropriate data and pressing "Save and Continue...species that the gene or insert sequence came from. Entering this information will populate a list of possible...

  29. Plasmids 101: Plasmid Incompatibility

    Type
    Blog Post
    ...system to partition themselves to each daughter cell during cell division. They don’t simply rely on chance...CBP), and a partition NTPase (Schumacher, 2012).  During cell division, CBPs bind to the centromere-like...replication that occurs for some types low copy plasmids during cell division simply does not allow for enough ...strategy has been successful in displacing, or “curing” the virulence plasmids from the bacterial pathogens... SR, Iredell JR (2017) Plasmid interference for curing antibiotic resistance plasmids in vivo. PLOS ONE...X, Wang D, Wang H, Feng E, Zhu L, Wang H (2012) Curing of Plasmid pXO1 from Bacillus anthracis Using Plasmid...22253811 Ni B, Du Z, Guo Z, Zhang Y, Yang R (2008) Curing of four different plasmids in Yersinia pestis using...

  30. Viral Production at Addgene

    Type
    Blog Post
    ...customers. Details about our production protocols, titering methods, and quality control are described below... or through collaboration with viral vector manufacturing facilities, such as the University of Pennsylvania...are then aliquoted and stored at -80 °C. Titer Titering is either performed by Addgene or by the University...University of Pennsylvania Vector Core. In general, titering is performed by the facility that produced the...those achieved using the standard qPCR method.  Titering by the University of Pennsylvania Vector Core ...Purity Purity of AAV preparations is assayed by comparing the relative stoichiometric ratios of the viral...aliquoted, frozen, and stored at -80 °C. Titer All titering is performed on lentiviral preparations that have...

  31. Using Video to Share Your Science: We Share Science

    Type
    Blog Post
    ...appeal, instant sharing across multiple other sites), I created a new platform for sharing video abstracts...University and developer of wesharescience.org. Sharing your research with the world can be challenging...printed journal articles as the dominant tool for sharing ideas. This leads to everyday obstacles for many...www.WeShareScience.org. WeShareScience is a free platform for sharing short, 3 to 5 minute videos about research. Videos...oceanographer who has a blog and several videos on sharing your research through video. Liz Haswell and ... 525.7569 (2015): 306-307. 3. Spicer, Scott. "Exploring video abstracts in science journals: An overview...Scientific Impact Data Freedom: The Expansion of Data Sharing in Research Publications Resources at Addgene....

  32. 5 Reasons to Use Reddit for Science Communication

    Type
    Blog Post
    ...the questions and preparing answers early in the day. Although I didn’t start answering until 5pm, frankly...well moderated. One of our original worries about sharing science advice on Reddit was that we’d be dragged... our AMA and Joanne’s preferred time to start answering questions. r/Science has pretty strict rules about...straight on through her designated two hours for answering questions. She was able to do this because we ...posted at 8:00 am and Joanne didn’t have to start answering until 5 pm). The ability to see the questions ... Tips: Online Social Networking for Scientists Sharing Your Lab Protocols: Using Apps to Save Time & Track...Experiments Additional Resources See How We're Sharing Science through Video with the Addgene Youtube ...

  33. Getting the Most from Your Lentiviral Transduction

    Type
    Blog Post
    ...culture has become overly confluent or too old. Ensuring that you are working with healthy, happy cells... the lentiviral stock needs to be titrated. If storing for less than a day, lentivirus can be kept at ...dry ice/ethanol bath or liquid nitrogen prior to storing. Know your target cells Labs tend to follow the...titer the virus (read this blog post for tips on titering your lentiviral prep!) and infect. While this ...consider is how the virus is being titered. Most titering approaches transduce into highly permissive cell...receptors, other important binding events take place during the course of infection. These binding events can...23589850. 9. Cronin, J., Zhang, X.Y., Reiser, J. “Altering the tropism of lentiviral vectors through pseudotyping...

  34. Celebrating Outstanding CRISPR/Cas9 Achievements at the Dr. Paul Janssen Award Dinner

    Type
    Blog Post
    ...acceptance addresses were inspiring! We also got to hear more from them during a panel discussion with ...bestowed upon discovers of CRISPR/Cas9 genome engineering, Dr. Jennifer Doudna and Dr. Emmanuelle Charpentier...the presence of scientific heroines is always inspiring. I was especially excited to attend this event...enough to make up for the somewhat naïve questions. During the panel, an artist was sketching her interpretation...accelerate the launch of clinical applications. During the panel discussion, Dr. Mello took it upon himself... in the room. He offered the great analogy of comparing the CRISPR/Cas9 system to Norton Antivirus. Antivirus...

  35. London Calling - Addgene’s New Europe Office in London

    Type
    Blog Post
    ...of Plasmid Sharing: Interested in more statistics from Addgene's 10 years of plasmid sharing? Check out...our US counterparts are still sleeping). Plasmid sharing in Europe: Addgene’s request statistics Since 2004...Developmental Physiology at the University of Paris VII. During my years in research, I developed several new plasmids...is excited to be working at Addgene, because it brings scientists together from all over the world to ...discussing a new scientific discovery or invention, and sharing those ideas with others. This love of interacting... blog post Or look at our "10 Years of Plasmid Sharing" Infographic!  ...

  36. Hot Plasmids: Summer 2024

    Type
    Blog Post
    ...structure determination LEA proteins protect samples during cryo-EM grid freezing Engineered and evolved optimizations...2024.01.21.576499.    LEA proteins protect fragile samples during cryo-EM grid plunge freezing By Rachel Leeson Cryo-EM...can be damaged at the air-water interface (AWI) during the freezing process, resulting in sample loss....protective methods or detergents like CHAPSO, while requiring a lower concentration of sample — nearly ten times...-water interface protectant for fragile samples during cryo-EM grid plunge freezing. bioRxiv 2024.02.06.579238...prime editing systems for specific applications! Delivering prime editors (PEs) via engineered virus-like...off-target editing. Cargo size limits are also less stringent for eVLPs than other viral delivery methods, allowing...

  37. Adeno-associated Viruses (AAVs) for Genome Editing

    Type
    Blog Post
    ... recently showed that by delivering CRISPR components as RNPs and delivering two halves of a repair template... Precision Medicine, CRISPR, and Genome Engineering. Springer, Cham, 2017. 29-42. PubMed PMID: 29130152... used AAV gene editing to introduce naturally occurring cancer-causing mutations into the endogenous allele...homology arms with the modification of interest, ordering them from a gene synthesis company such as IDT...anaphase, and chromosome segregation in human cells harboring tumor-derived mutations in STAG2." PLoS genetics...Howes, Rob, and Christine Schofield. "Genome engineering using adeno-associated virus (AAV)." Chromosomal...

  38. Transferable Skills Guide: Time Management

    Type
    Blog Post
    ...goals are (see monitoring task progress), and I know that I’ve blocked out time during my day according...transferable skills while working in the lab and by offering advice on areas where you might need some help... can spend accomplishing small tasks (like... preparing plasmids to send to Addgene). Following my advice...too many activities too close to one another. Monitoring task progress. For this, I always make sure I...-world example, let’s say I’ve blocked out time during my day to write a blog post, edit other blog posts...leave your phone in your bag on “Do Not Disturb” during the times you’ve blocked out for other activities...instance, while I may have found splitting cells boring, hanging out in the tissue culture room also gave...

  39. Hot Biosensors 2022: Year-End Roundup

    Type
    Blog Post
    ... for monitoring RTK signaling.  Figure 4: Design of pYtag fluorescent biosensors for monitoring RTK activation...for investigating neuronal signaling and for engineering improved sensors for these and other targets....class of signaling receptors, but the options for measuring activity of specific RTKs are limited. In a new...colleagues describe pYtags, a modular system for monitoring specific RTK activation in vivo (Farahani et ...sensors. Using genome mining and structure-guided engineering of potassium binding proteins, they developed...and are a promising starting point for further engineering. Figure 5: Structure-guided design and genome...s41467-022-30685-x. Feel the heat with B-gTEMP  Measuring local temperature variations and heat transfer...

  40. Plasmids 101: Common Lab E. coli Strains

    Type
    Blog Post
    ...of animals. There are many different naturally occurring strains of E. coli, some of which are deadly to...strain is a bit more convoluted due to researchers sharing and renaming samples throughout history. It was...cloning, routine cloning, cloning unstable DNA, preparing unmethylated DNA, and more. Many mutations that...Consequence dam DNA adenine methylase mutation (GATC) Preparing unmethylated DNA, important when trying to cut... dcm DNA cytosine methylase mutation (CCWGG) Preparing unmethylated DNA, important when trying to cut...relA1 Relaxed phenotype, mutation eliminating stringent factor Allows RNA synthesis in absence of protein...lacZΔM15] hsdR17(rK-mK+) JM110  Streptomycin  For storing plasmids that should not be Dam or Dcm methylated...
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