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TypeBlog PostPublishedSept. 1, 2015, 2:30 p.m....start defining keywords you’ll want to find in candidates’ resumes or CVs; “vivarium”, “weaning”, “colony...developing a budget, I suggest that you focus on defining these resources and not on the financial aspects...your current P.I., lab manager, or administrator containing all of the materials that you’ve used within ...colony”, etc. This will help save time by screening out irrelevant candidates, and, more importantly, it...Channel your inner curiosity and use your research training to help you find other solutions to reach your...
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Synthetic Biology, Artificial Chromosomes, iGEM & More - Interview with Tom Ellis
TypeBlog PostPublishedNov. 29, 2017, 2 p.m....advice for anyone pursuing a career in academia. Learning how to build a lab and a research program and ... biology to the UK with foundational research, training, startups, and real applications of SynBio. Chapter...seen with things like DNA synthesis, Golden Gate cloning, and CRISPR. The yeast synthetic genome project... of Cambridge before spending time in biotech screening direct interactions between drugs and promoters...Nonprofit BioBuilder Plasmids 101: Golden Gate Cloning Learn About Working at a SynBio Startup in Our ... -
Generating Mouse Models Using CRISPR/Cas9
TypeBlog PostPublishedJuly 12, 2016, 2:30 p.m....Many thanks to our guest bloggers, Wenning Qin and Haoyi Wang! Wenning Qin is a molecular biologist with.... This post was contributed by guest bloggers, Wenning Qin and Haoyi Wang. CRISPR/Cas9 is revolutionizing...al., 2013). Like everyone else in the world, I (Wenning) was fascinated by CRISPR and rushed to test it... hard. I was mentally prepared to go through a learning curve that would result in success only after ...broken ends together through non-homologous end joining (NHEJ), this will leave behind a “scar”, with nucleotides... -
How to Design Your gRNA for CRISPR Genome Editing
TypeBlog PostPublishedSept. 24, 2020, 1:15 p.m.... involves the need for single cell cloning and subsequent screening for successful edits. This is a very...important to remember that the same is true when designing gRNAs for using CRISPR technology – the “best”...and sequence are important considerations for designing your gRNAs. For indels, it's not so important ..., the error-prone nature of non-homologous end joining (NHEJ) often leads to the generation of indels ...requires not one but two rounds of single cell cloning – as a control, one should revert the edit back... -
Microbes: Look, Listen, and Tell
TypeBlog PostPublishedAug. 28, 2018, 12:04 p.m....may think about sitting amongst a sea of chairs listening to talks all day. But nope, not at the American...everyone - everywhere! From ‘looking at’ to ‘listening to’ microbes But seeing microbes through a microscope...Shapiro from Caltech presented an exciting way of listening to microbes by using non-invasive methods for ...approaches to presenting the microbial world After learning about using sound to visualize bacteria in the...Raymond W., et al. "Acoustic reporter genes for noninvasive imaging of microorganisms in mammalian hosts.... -
What's New in CRISPR - December 2019
TypeBlog PostPublishedDec. 17, 2019, 2:37 p.m....Synthetic Biology Find the CRISPRi toolkit SECURE adenine base editor Earlier this year, Keith Joung’s lab...to rat APOBEC1. Now, they’ve engineered SECURE adenine base editor variants using a protein-truncation...pyogenes Cas9. They called this editor Target-ACE (adenine and cytosine editor). Target-ACE exhibits increased...inactivated RNA-targeting Cas13 (dCas13) with the adenine deaminase domain of ADAR2. Now, they’ve further...this editor for C-to-U editing by evolving the adenine deaminase domain of ADAR2 into a cytidine deaminase... -
Modulate the Activity of 17 Signaling Pathways with One Kit!
TypeBlog PostPublishedMarch 22, 2016, 2:30 p.m....colleagues at Duke University, in collaboration with “cloning guru” Kathleen Ottina and David Sabatini at the...which is now available at Addgene. “We called it cloning by mail,” Wood says, and it took them about six...and his colleagues developed the kit for use in screening for pathways that drive resistance to drugs in...Tracing Read How CRISPR Can Be Used in Genome-Wide Screening Browse 28 Hot Plasmid Technologies from 2015 ... cancer screening... -
X-CHIME: Context Dependent Germline Knockout in Immune Cells
TypeBlog PostPublishedJan. 16, 2024, 6:13 p.m.... G., Haining, W. N., & Sharpe, A. H. (2019). A CRISPR-Cas9 delivery system for in vivo screening of genes...hematopoietic stem cells with a lentiviral vector containing a gRNA. These transduced stem cells can be implanted...Huang, A. Y., Anekal, P. V., Montero Llopis, P., Haining, W. N., Doench, J. G., & Sharpe, A. H. (2024). ..., Gaudiano, E.F., Al Abosy, R., Freeman, G.J., Haining, W.N., & Sharpe, A.H. (2019). PTPN2 regulates the... -
Easing the Protein Purification Process with pCri
TypeBlog PostPublishedJune 19, 2015, 3:08 p.m....to improve protein stability and solubility. Determining the best way to express one’s protein of interest...specialists recommend doing some research, notably determining the subcellular localization and the number of...plasmids uses the same restriction enzymes for cloning (see schematic below), enabling a researcher to... in E. coli, the use of a thioredoxin A (TRX)-containing vector can improve disulfide bond formation. ...tags Resources on Addgene.org Find Additional Cloning Kits ... -
CRISPR Protocol for Genomic Deletions in Mammalian Cell Lines [Video]
TypeBlog PostPublishedFeb. 18, 2015, 3:09 p.m....finger nucleases to harness non-homologous end joining repair (NHEJ) [1]. Our initial work was with TALENs...haploinsufficiency. Subsequent modifications at the remaining allele within the deletion boundaries may be produced... paired DSBs. This method allows for deletion screening by conventional PCR. Biallelically disrupted cells...transfected cells via FACS, primer validation, and screening by PCR for biallelic deletion clones: ...paper describes their methods for CRISPR design, cloning, and delivery, as well as their techniques for ... -
A History of Genome Engineering in Popular Culture
TypeBlog PostPublishedFeb. 25, 2020, 2:15 p.m....simulations and machine learning to develop a predictive understanding of the adenine base editors which can... regulating and propagating life, there was no turning back. While the world of molecular genetics was...This is typically followed by non-homologous end-joining that results in gene knockouts. Star Trek: Into...locations in the genome. This was soon followed by the adenine base editor (ABE) enzyme in 2017. ABEs rely on ...editor proteins. She is particularly interested in combining the realms of art and science. Follow her at @... -
Tips for a 1st Time CRISPR User (by a 1st Time CRISPR User)
TypeBlog PostPublishedMarch 7, 2017, 3:30 p.m....randomly chose one of my stable pools. Viruses containing BRAF gRNA (BRDN0000561167 and BRDN0001146266)... 4). The most difficult part of the assay was designing primers flanking my edits in genomic DNA and doing...gRNA-transduced samples (data not shown). Pro-Tips: 1.Designing primers against genomic DNA to amplify the gRNA... vectors and genome-wide libraries for CRISPR screening. Sanjana NE, Shalem O, Zhang F. Nat Methods. 2014...Additional Resources on the Addgene Blog Genome-wide Screening Using CRISPR/Cas9 Deliver Multiple gRNAs at Once...Lentivirus Production Websites Primer3: for Designing Primers UCSC Genome Browser: For eploring genomic... -
Finding Your Science Policy Path
TypeBlog PostPublishedSept. 22, 2020, 1:15 p.m....field from a more traditional policy background, learning the science part on the job. Each path to science...implementing the policy within the jurisdiction of the governing body. This type of work can happen at the national... professional societies, and industry through convening activities, formal lobbying, or strategy setting...The “little p” policy is non-governmental work happening at large organizations who have to develop policies... do you do science policy? Many science policy training programs and fellowships have a strong focus on...nation’s largest integrated healthcare system? Defining your why is what will make you a competitive candidate... -
Live and Let Dye: Self-Labeling Protein Tags
TypeBlog PostPublishedApril 16, 2024, 1:15 p.m....you’ll probably find lots of different plasmids containing the same protein of interest tagged with different...provide many advantages over fluorescent proteins, opening up new applications. Figure 1: Live-cell...covalently linked to derivatives of its substrate O6-benzylguanine (BG) (Keppler et al., 2003). This engineered...FlAsH, ReAsH) Few Easy SNAP-tag 182 Benzylguanine (BG) Many Easy eDHFR 159 Trimpethoprim...Bottanelli et al., 2016) under CC-BY license. Combining orthogonal self-labeling tags on different proteins...able to get a plasmid ready-to-use without any cloning at all! As mentioned above, several variants of... -
A Guide to Getting Started in Undergrad Research
TypeBlog PostPublishedJune 8, 2021, 1:15 p.m....Getting more scientific and professional training after earning a doctorate Considered a "stepping stone...also more than eager to help you along your path. Learning about their career paths and goals helped me reflect...research opportunities are also fantastic for broadening your experiences! These applications can be highly...years. But college, and science, are places for learning and growth! As you learn more about yourself and...EleanorWang. References and resources References Canning EA, LaCosse J, Kroeper KM, Murphy MC (2019) Feeling... -
What's the Best Way to Elute and Store Your Plasmid DNA?
TypeBlog PostPublishedJune 4, 2020, 1:22 p.m....glycosidic bond leads to the release of a purine (adenine or guanine) from the DNA. These apurinic sites if in...used for plasmid amplification are endA negative, meaning the gene has been removed/mutated to neutralize...without a kit Read our blog posts about plasmid cloning Browse our Plasmids 101 blog posts Resources on...Molecular Biology Reference Find protocols on plasmid cloning Find plasmids for your research ... -
Addgene-Seeding Labs Plasmid Grant: Accelerating Science Globally - 5 Scientists, 4 Countries & 3 Continents!
TypeBlog PostPublishedJuly 18, 2017, 12:41 p.m....grant this year. These scientists work on projects spanning from the identification of plant compounds and...activities (1). She has developed a fast endophyte screening system and plans to employ the light‐inducible...integration vectors (see for example this yeast cloning kit) she intends to make yeast capable of desaturating... to bacteria and plants. Her research involves cloning cry genes from the soil bacterium Bacillus thuringiensis...bacterial expression system such as the pET GFP LIC cloning vector and the pOKD plasmids. The recombinant Cry... -
10 Ways to Share Your Science!
TypeBlog PostPublishedMay 13, 2014, 8:34 p.m....Morehead Planetarium. Similar things are likely happening in your community, and, if they aren’t, think ...motivated more by the desire to help close the widening gap between science and society by connecting ...out how they got where they are. 8) Look into training from an organization such as COMPASS. Although...their science matter. If you aren’t ready for a training or one isn’t available yet, order a copy of Nancy...University of California, Santa Cruz, has been turning scientists into confident and connected science... -
History of CRISPR Cas - A tale of survival and evolution
TypeBlog PostPublishedNov. 25, 2013, 10:12 p.m....Horvath P, Moineu S, Mojica FJM, Wolf YI, Yakunin JvdO, Koonin EV. Nature Reviews Microbiology. 2011 Jun...powerful and adaptive defense strategy against life-threatening viruses. Evolving to survive Although the CRISPR...CRISPR/Cas adaptive immune system seems like the winning ticket to ensure prokaryotic survival; however,...Cong et al. and Mali et al. in January 2013 (7, 8). Nine months later, more than 1500 publications describe... -
Science Careers: Unruly Interests Feed Many Paths
TypeBlog PostPublishedSept. 26, 2017, 12:12 p.m..... My eye was caught by a couple of unusual job openings, although they seemed too far afield at first ...forget: what would it be like? Then I spotted an opening for being an editor at Science magazine. That one...referees. And I am constantly teaching – often explaining a technical piece of research to others outside...of the world’s scientific discoveries into an entertaining podcast? Look at our multimedia producer and...Blog Posts Resources at Addgene.org Find Job Openings at Addgene ...