We narrowed to 845 results for: CAL;
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TypeBlog PostPublishedDec. 8, 2016, 3:30 p.m....developed cloning vectors to harness it. These vectors, called pKIL18 and pKIL19, contained the ccdB gene in frame...ccdB and therefore would die. This procedure dramatically reduces the number of clones that do not contain...and ccdB Survival strains have been developed specifically with ccdB-containing vectors in mind, any plasmid...
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The Breast Cancer Microenvironment: A Tumor’s Backstage Team
TypeBlog PostPublishedJan. 31, 2019, 1:32 p.m....reprograming influence tumors development? Being pathological active regions, microenvironments shape a tumor...interested in science journalism and writing about biomedical issues. Follow her on Twitter @BrbaraPinho6. ...expression predicts early tumor recurrence and poor clinical outcome in human breast cancers." The American... -
Human Germline Editing Using CRISPR
TypeBlog PostPublishedAug. 10, 2017, 2:19 p.m.... rate would need to be much, much lower before clinical applications could be considered. It is important... efficiency and applicability. Given moral and ethical concerns surrounding such research, it is crucial... experiments are needed before we can consider clinical applications for germline editing. Although this... -
Writing Scientific Manuscripts: Literature Searching, Reading, & Organizing
TypeBlog PostPublishedMay 5, 2015, 3:54 p.m.... year, narrow by article type (review, patent, clinical study, research article etc). There are also numerous...conclusions. In any case, the reader should give a critical eye to all results in the relation to how they... biochemistry instructor in the Department of Biological Sciences at Louisiana State University. She is... -
A Look Back at One Year of Plasmid Sharing for COVID-19 Research
TypeBlog PostPublishedJuly 12, 2021, 1:15 p.m....., 2021). Another version of the spike protein, called HexoPro, was developed by Jason McLellan’s lab....genome-wide CRISPR libraries, Brunello (knock out) and Calabrese (activation), from Addgene to better understand...bidirectional CRISPR screens identify mucins as critical host factors modulating SARS-CoV-2 infection. ... -
Lab Automation at Addgene
TypeBlog PostPublishedMarch 22, 2018, 1:28 p.m....bacteria through the mail due to laws concerning genetically modified organisms. Since Addgene is very proud...individually. Enter the BIOROBOT! This robot is called the Qiagen BIOROBOT Universal Platform and it suits... for a mini prep, except it does it on a larger scale. Using the BIOROBOT Universal Platform is ideal ... -
Meet an Addgenie: Insights into Grad School, Career Paths, and More
TypeBlog PostPublishedJune 25, 2020, 1:15 p.m....Addgene’s Lab Manager, was taking classes at her local community college when she took her first job at... and hoping to pursue. The choice to not go to medical school was devastating. At the same time, she has...circumstance. She was working as a research and technical assistant, when her life changed––she had a daughter... -
CRISPR Activators: A Comparison Between dCas9-VP64, SAM, SunTag, VPR, and More!
TypeBlog PostPublishedOct. 6, 2020, 1:15 p.m.... to be re-engineered for each gene, making wide-scale gene activation seem next to impossible. The development...methods greatly improved CRISPRa, expanding its practicality and popularity in research (Tanenbaum et al....JS, Nishimasu H, Nureki O, Zhang F (2014) Genome-scale transcriptional activation by an engineered CRISPR-Cas9... -
6 Tips for Grant Writing
TypeBlog PostPublishedOct. 18, 2016, 2:30 p.m....unintelligible to a lay audience. My lay paragraph typically includes a couple of sentences of introduction... rejected on some occasions. Strategize, choose calls that suit your career stage / research area, and...peers and collaborators can also help you get some critical insight into what you're doing wrong. All this... -
Reaching out to China: Canton Nucleic Acids Forum (CNAF) 2015
TypeBlog PostPublishedDec. 1, 2015, 3:30 p.m....remarkable progress on establishing a system for the chemical replication of RNA in the absence of evolved enzymatic... about using CRISPR/cas to address the growing clinical problem of antibiotic resistant bacteria. See ... and Sun Yat-sen University presented two new technical developments. Using the Tet-on system, his lab... -
xCas9: Engineering a CRISPR Variant with PAM Flexibility
TypeBlog PostPublishedMarch 28, 2018, 6:52 p.m....edits or knock-ins. Editing efficiency decreases drastically once your cut site is more than 10 bp away from... mutagenesis plasmid (MP6) permits quick, large-scale mutation of the gene of interest. They created a... a selection phage (SP) to carry catalytically dead SpCas9 (dCas9) fused to the bacterial polymerase subunit... -
Multiplexed Capture of Promoter-enhancer 3D Chromatin Structures Using CRISPR
TypeBlog PostPublishedJuly 2, 2020, 12:40 p.m....but close together in terms of where they are physically located. DNA is then sheared into pieces small...on-target enrichment. 2. Determining the spatial and hierarchical organization of chromatin interaction networks...CAPTURE 2.0 is the construction of spatial and hierarchical chromatin interaction networks. Think of these... -
CrispyCrunch: High-throughput Design and Analysis of CRISPR+HDR Experiments
TypeBlog PostPublishedFeb. 7, 2019, 2:16 p.m....sequencing primers for quality control. At such a scale, automation is crucial, both for speeding things...to the same principles listed here. It employs biological best practices. For example, it ranks guides ...published on how to do it best. Scientists are typically advised to mutate the PAM to which the guide binds... -
Plasmids 101: Multicistronic Vectors
TypeBlog PostPublishedSept. 9, 2014, 8:20 p.m....bicistronic vectors take advantage of an element called an Internal Ribosome Entry Site (IRES) to allow...use the 2A peptides instead of an IRES when stoichiometrically equivalent levels of expression are required...journal.pone.0018556. Epub 2011 Apr 29.) PubMed. 5. Scalable signaling mediated by T cell antigen receptor-... -
An Introduction to Adenovirus
TypeBlog PostPublishedOct. 3, 2023, 1:15 p.m....culprit for these unusual cases was either 1) a physiological effect of SARS-CoV-2 toward destabilizing one...Adenovirus (and/or to its partner, AAV) or 2) a sociological effect of the COVID-19 lockdowns causing a lack...Adenoviruses: General Features. In Reference Module in Biomedical Sciences. Elsevier. https://doi.org/10.1016/B978... -
CRISPR 101: Making a Knock-In Cell Line
TypeBlog PostPublishedDec. 22, 2022, 2:15 p.m....not the case for knock-ins. Knock-in mutations typically require an exact DNA sequence to be incorporated...know that HR is often mutated in cancer cells, specifically in breast and ovarian lines commonly used in...common editing pathway for DNA repair. To tilt the scales in your favor for an HDR edit, you can try one ... -
DNA Purification Without a Kit
TypeBlog PostPublishedFeb. 11, 2020, 2:15 p.m....methods avoid many of the drawbacks of kits and typically follow the same principles: cells are lysed, RNAs...method described in Kim and Morrison is nearly identical to that of commercially available kits but instead...B., & Gillespie, D.A. (1979). Preparative and analytical purification of DNA from agarose. Proceedings... -
Simplify Cloning with in vivo Assembly
TypeBlog PostPublishedOct. 18, 2018, 12:37 p.m....-55 °C for the homologous region (calculated using Oligo Calculator). Insertions - To insert short sequences...parental DNA (37 °C for 15 mins). This enzyme specifically cuts methylated DNA, so it will selectively ... -
CRISPR 101: Ribonucleoprotein (RNP) Delivery
TypeBlog PostPublishedSept. 6, 2018, 12:02 p.m....Liang et al. 2017). PEG-mediated methods, while technically less difficult to perform, often show low efficiency...ribonucleoprotein electroporation of zygotes." Journal of Biological Chemistry (2016): jbc-M116. PubMed PMID: 27151215...Cell-Type-Specific Gene Editing." Journal of the American Chemical Society 140.21 (2018): 6596-6603. PubMed PMID:... -
The Importance of a Fun Workplace: Company Culture at Addgene
TypeBlog PostPublishedSept. 3, 2015, 2:30 p.m.... familial aspect of Addgene. As our size has drastically increased, it would have been a tight squeeze...event! Addgene company culture At Addgene we organically developed the philosophy of “work hard, play...department) right in your breakroom. 5. Throw some physical activity into the mix It’s easy to sit around ... -
Negotiating Work and Life: How to Find the Joy
TypeBlog PostPublishedMarch 20, 2014, 3:04 p.m....community to help Involve family. If you don’t have local family to help, create “fake” family by forming ...help you do your job better. #8 Thou shalt enthusiastically foster independence Your partner can make ... fold their laundry, take public transportation, call a cab and remember their own school assignments,... -
Tips for Writing a Good Cover Letter
TypeBlog PostPublishedMay 21, 2024, 1:30 p.m....writing are key skills. Other fields, often highly technical or skills-based ones, may not use them very often...questions about your application. These could be logistical, like briefly explaining a gap in your job history...letter needs to be coherent and free of typos and grammatical errors. It does not need to be a literary masterpiece... -
Antibodies 101: Reading a Flow Plot
TypeBlog PostPublishedFeb. 6, 2024, 3:13 p.m....generate a dot plot (Figure 1). Clustered events are called a population. ...of markers you use to define your population is called your “gating strategy.” This series of markers ...around clusters to define populations. This shape is called a ‘gate’; it tells the machine that everything ... -
Viral Production at Addgene
TypeBlog PostPublishedOct. 24, 2023, 1:15 p.m.... relative to empty particles (black arrowhead). Scale bar = 100 nm. AAV resources Browse AAV preps... a concentration step prior to being frozen. Specifically, lentiviral particles are collected from the... strain of E. coli. In addition, plasmids are typically prepared using endotoxin-free plasmid purification... -
How to Write a Scientific Review Article
TypeBlog PostPublishedFeb. 16, 2017, 3:30 p.m.... to synthesize a large volume of information: basically, a scientist’s tool kit. I wrote a review article... was cheating. Eventually I learned that it’s practical because there simply isn’t enough time to read...used both. Curate and present some useful data Typically lists and pictures are the most useful parts of... -
Four Base Editing Reporters to Monitor and Enrich Editing in Real-time
TypeBlog PostPublishedJuly 7, 2020, 1:15 p.m....are subject to excision and ssDNA cleavage by canonical base excision repair enzymes. At the same time...targeted by a cytidine deaminase base DNA editor. Specifically the BFP mutant (BFPH66) contains a histidine...markers, and other enzymes such as Cre-recombinase drastically expanding the base-editing reporter toolbox. ... -
Binning Singletons: Tackling Conference Networking When You Don’t Know Anyone
TypeBlog PostPublishedFeb. 18, 2020, 2:15 p.m....by Joe James from Binning Singletons. The sheer scale of a large conference can be intimidating. And it...when we bring all the bins together at a mixer. We call this Quorum Sensing, after the phenomenon in bacteria... career scientists, students, and those from historically underrepresented groups. Through such mentoring... -
Save Time with Transient Plant Leaf Transformations
TypeBlog PostPublishedJuly 25, 2019, 12:03 p.m....below. The GUS gene (β-glucuronidase) with histochemical GUS-staining is a very useful visual reporter...protein of interest to elucidate the subcellular localization of your protein of interest. Find all plasmids...see Figure 3). This method is probably the most scalable method. Transfer your plant into the dark 16 ... -
Overcoming the AAV Size Limitation for CRISPR Delivery
TypeBlog PostPublishedSept. 16, 2020, 1 p.m....SpCas9 and SaCas9, Zhang’s group used an approach called BLESS (direct in situ breaks labeling, enrichment...post-injection. Livers from these mice were histologically normal and liver injury markers were not increased...enough to deliver with AAV. This system fuses catalytically dead dCas13b to the catalytic domain of RNA ... -
The PAM Requirement and Expanding CRISPR Beyond SpCas9
TypeBlog PostPublishedAug. 20, 2020, 2:30 p.m.... D1135E variant is far more selective for the canonical S. pyogenes PAM sequence NGG compared to wild-...novel CRISPR proteins has and will continue to dramatically increase the number of CRISPR applications. ...Wolfe SA, Sontheimer EJ (2018) NmeCas9 is an intrinsically high-fidelity genome-editing platform. Genome...