...   Some phage display libraries screen antibody genes from B cells of immunized animals, but it's possible...screening scFv generated from the B cell antibody genes from diseased or vaccinated humans. How do you produce...cytoplasm. expression in the presence of molecular chaperones that help scFvs fold properly in the cytoplasm...also be fused to cellular toxins, radioisotopes, cytokines, and enzymes for cancer, autoimmune, and/or inflammatory...

...demonstrate a POI’s absence. It also helps confirm that lanes are loaded evenly. Created with BioRender.com. ...blot. So how can you accurately compare different lanes? In relative quantification, the POI bands are normalized...blue) to infer or calculate things about earlier ones. In practice, multiple sources of error (red) influence...detection range of the assay is between the two dotted lines. Created with BioRender.com. To see why this...

...powerful site-specific knock-in tool that can be harnessed by researchers for many applications. Your favorite... chromosomal aberrations. Pfeiffer P, et al. Mutagenesis (2000). PubMed. Multiple Pathways of Recombination... Increased Efficiency of CRISPR/Cas-mediated Transgenesis (2018). Pubmed. The Road Less Traveled: Strategies... Increased Efficiency of CRISPR/Cas-mediated Transgenesis (2018). Pubmed. CRISPR References: Genome ...

...promoter-luciferase constructs. In multiple HIV-1 latent T cell lines, as well as a latent microglial line, treatment ...via HIV-1 promoter-driven GFP expression. In cell lines that can produce HIV-1 toxic proteins, CRISPR/Cas9...Cas9 have also been used to create mutations in genes essential to HIV-1 replication. However, this approach...modification targeting either the LTRs or essential genes. Many of these escape mutations were located near...

... Cre-dependent transgenes can also be paired with Cre-expressing transgenic mouse lines to restrict transgene...used and emerging capsids to help you decide which ones may be suitable for your experiment. Like all viral...inducing an immune response against the capsids or transgenes. Sensory neurons and liver appear especially ...appropriate cell type-specific promoters or Cre lines. Spatial transcriptomics suggests that AAV-PHP.eB... Seed, C., & Deverman, B. E. (2019). Delivering genes across the blood-brain barrier: LY6A, a novel cellular...

...them. There are suspected cases of insertional mutagenesis in humans (1) and AAVs are non-enveloped viruses... estate mantra, the choice of serotype (which determines which tissues and cells an AAV can infect) can... Russell, David W. "AAV vectors, insertional mutagenesis, and cancer." Molecular Therapy 15.10 (2007):... PubMed Central PMCID: PMC3098627. 6. Salegio, Ernesto A., et al. "Axonal transport of adeno-associated...

...decision. If I’d spent my senior year isolating pheromones from various C. elegans mutants by myself, I ...etc. If you find that there is not someone (or someones) taking on these responsibilities on a team to...really starts functioning (or malfunctioning). Clear lines of communication Few people actually like meetings...purpose, an excellent leader, and perfectly clear lines of communication, but if teammates don’t trust each...

...methods. However, most investigators use single cell clones for in vitro CRISPR experiments. The likelihood...may not be worth the cost and labor when single clones are selected. Conversely, translational research..., Yanfang, et al. "High-frequency off-target mutagenesis induced by CRISPR-Cas nucleases in human cells...mutations triggered by engineered nucleases." G3: Genes| Genomes| Genetics 5.3 (2015): 407-415. PubMed PMID...

...1 will get you in the ballpark for average size genes and vectors, this ratio is really meant to refer...

... analysis, neighborhood analysis of surrounding genes, and experimental data. In some cases, CRISPR systems...researchers began building optimal plasmids to harness the power of Class 1 systems. Class 1 CRISPR systems...cas10. All type IV subtypes are missing adaptation genes, Cas1 and Cas2, which are critical for inserting...A–C, with the beloved SpCas9 (from Streptococcus pyogenes) belonging in II-A. Cas9 was the first to be adapted...

..., energy is propagated into the sample via an evanescent wave that only excites fluorophores within a ...through the fluorophore’s interaction with an evanescent energy field. Lightsheet microscopes that project...sheets of excitation light through multiple focal planes of a sample may also be useful for providing fast...Protein Resources Find Empty Fluorescent Protein Backbones Find Plasmids for Optogenetics ...

...protein, you can site-specifically label it with finesse using tRNA suppression technology (Fig. 1) (Dumas...cells nonspecifically, and they cannot cross cell membranes, so they are best used either with plasma membrane... Most importantly, these compounds cross cell membranes, thus enabling FRET measurements in intact cells...Fessenden is currently an Assistant Professor in the Anesthesia Research department at Brigham and Women’s Hospital...

...sequence between two target sites. Modifying multiple genes at once. Using multiplexed gRNAs can target multiple...CRMAGE system is a fast, multiplexable method that combines CRISPR and recombineering-based MAGE (Multiplex...simultaneous multiplex editing of six different genes. This system doesn’t rely on any additional or host-dependent.... Yin, Linlin, et al. “Multiplex Conditional Mutagenesis Using Transgenic Expression of Cas9 and sgRNAs...

... experience and judgement to a wide variety of business decisions that are not “legal” at all. Q: How ...Addgene board member? BN: Having seen so much in the business world from the mindset of a lawyer can bring a...if we do this, then what?” I hope to bring that business experience and judgement to the organization and...

...proteomic tools to discover genes and proteins associated with pathogenesis. In his future work. Dr. Bengyella...CRISPR-based gene repression tools to find and test genes that may play a role in the development of uterine...

...science and business or programming were new to some, who were surprised at “how much business there is ...Last week, 20 students and four chaperones from Denis-Yarmouth Regional High School visited our labs ...

...that’s considered unlikely to change – but the illness has been fatal in 70 percent of the most recent...infection also makes it a critical component of vaccines and the target of neutralizing antibodies and ...Ebola representing the latest outbreak and earlier ones, along with links to the relevant scientific literature...

...writing, clinical, regulatory, product development, business development, consulting, policy, big data and ...medicine and health, education, law, accounting, business specialties, theology, and the arts, and requiring...your work has impacted your field (i.e. finding vaccines for deadly disease, building algorithms to program...regulatory, project management, alliance management, business development, finance, and operations. To learn...

...My son is an engineer and my daughter has a business degree. From the very start of their undergraduate...of driving systemic change in academic career readiness training. It's been an honor to work with such... essential that we integrate inclusive career readiness training in this educational path before we lose...

...to knock-out single genes in clonal populations and to knock-out thousands of genes in mixed population...targeting over 19,000 and 20,000 human and mouse genes respectively, use lentiCRISPR v2 as the backbone...

...the CRISPR/Cas9 system to precisely edit specific genes in monkeys. "Our study shows that the CRISPR/Cas9...system enables simultaneous disruption of two target genes in one step without producing off-target mutations...the news on CRISPR has gotten, consider these headlines in The New York Times and Forbes respectively:...

...the stage arm to move the stage so that the light shines onto your sample. To ensure that your sample is...

... the biomedical research community. Along these lines, Darrell recently joined the Addgene Board of Directors... various lung diseases. Chapter 10: How Darrel defines Open Science and how he practices and promotes ...open science in the lab - sharing reagents, cell lines, data, and knowledge without exclusivity. Chapter...

...shared resources. As we step into 2025, these milestones remind us of the extraordinary impact that happens...in line with current Web Content Accessibility Guidelines (WCAG). These changes mean our content is both... Chonnettia Jones...

...culture of engagement in the issues at hand and eagerness to get, as well as give, information to make improvements...to the manager. Team members rarely rush to give honest feedback to the manager for a variety of reasons... frequency. Anonymity can be a great way to get honest feedback, but shouldn't be the only way.   From...

...perform pooled CRISPRi screens to identify essential genes and phage host factors, and identify sequence-specific...Knockout Library is a lentiviral library targeting 26 genes chosen from an analysis of human metastasis cohorts...knockout library targeting mouse tumor suppressor genes (TSG). Find the Mouse Metastasis CRISPR-Cpf1 Double...

... more readily enable research. Often “normal” timelines that could be weeks to months were reduced to ...Institute                                       Chinese Academy of Medical Sciences University of Texas...Stanford University University of Iowa University of Minnesota See highlights from the COVID-19 collection Partnership...

... cell’s surface proteins and this interaction determines the specificity and efficiency of the AAV serotype...each approach and to follow best practices and guidelines. It was wonderful to see the development of this...by the workshop participants  - a testament to openness and sharing in science.  We’re excited to support...

...Targeted manipulation Controlling stimulation Invasiveness  Choosing the best set of tools therefore depends...precise and the offset of stimulation is gradual. Invasiveness Both optogenetic and chemogenetic tools need...region Impacts all cells expressing receptors Invasiveness Mice require permanent intracranial implant ...

... may be electrochemical or light-based, with luminescent and fluorescent reporters being especially popular...characteristics like pH, redox state, and voltage across membranes. These sensors generally follow the same pattern...endoplasmic reticulum (ER), vacuole or cellular membranes. Examples include blue flame mitochondria-targeted...

...able to obtain eight clones. We ran digested and undigested samples of the clones on an electrophoresis... and I were unsure if there were any successful clones because the bands were all in the wrong place. ...

...easy to spot meeting participants. They were the ones holding poster tubes (or as our baffled flight attendant...specifically in Dr. Zhang's introduction. His willingness to share reagents so freely with the academic...changing it from a puzzle to decrypt to a tool to be harnessed. Sharing CRISPR, synbio, & more via Addgene Addgene's...

...it takes to be a good scientist are not the same ones it takes to be a competent manager (much less a ...strong sense of engagement in the work Behind-the-scenes support from their bosses and employers. Availability...may not necessarily be comfortable with imposed deadlines (there’s that need for flexibility, sometimes ...

...Vectors (or empty backbones) are frequently used in molecular biology to isolate, multiply, or express...At Addgene, we have a vast collection of empty backbones that have been designed, tested, and published...vector). Check out our complete guide to empty backbones for additional details and guidance for choosing...

...discards according to the Cambridge, Massachusetts guidelines and wear an extra sweater so we don’t have to...visualization or isolation, then using only a few lanes or a small portion of the gel.  What a waste?  I... pour over the sides.  These tiny gels and tiny lanes required less agar, less ethidium bromide, less ...

...Tuesday (120 h post-transfection): Add 4% BCD Feed. Wednesday (144 h post-transfection): Add 4% BCD Feed. Thursday...

...20th century, and many iconic papers, including the ones on the structure of the DNA, were not peer-reviewed...submitted paper in addition to the journal-specific guidelines.  Peer-review step 1: The first read At the first...preferably the following morning - as usually deadlines are pressing. During this second read, I pay more...

...sometimes impossible) timelines—others are usually waiting Work is concentrated to business hours (mostly), ...with peer mentors, to discuss your strengths, weaknesses, interests and passions is another great way ...

...communication itself, not the communication method, that determines if something is scientific or science communication...defined by any number of characteristics. Common ones include age range, familiarity with the subject,...audience to engage if they so wish. (And let’s be honest: are you more likely to closely read an email sent...

...fall into one of two categories: ones induced by the researcher and ones that happen naturally.  Sample ...working with native proteins instead of denatured ones - well, don’t relax just yet. Proteins often change...