...transcription factors, starting with an established method called ChIP-seq. ChIP-seq relies on antibodies to recognize...We’ve been doing that for years using this assay called ChIP-seq, but it’s really hard for lots of transcription... the second plasmid inserts a small epitope tag called FLAG at the end of the transcription factor. It...have good antibodies. The results look almost identical. Addgene: What advantages does your system have...happy to provide them, that never works on a large scale. The world is too big now. Mendenhall: So many groups...

...missing or damaged nucleotides that are common to biological sources of DSBs, and which need to be processed...generated in the course of repair by NHEJ are typically small (1-10 bp) but extremely heterogeneous. There.... NHEJ can also be engaged by variants of the canonical Cas9 approach. A pair of CRISPR guides that flank...Translocations in Human Cells Are Generated by Canonical Nonhomologous End-Joining. Molecular Cell 55:829...CRISPR Libraries Offer Genome-Wide Control for Large-Scale Functional Screens        ...

...ribonucleoprotein, and AAV.  Read the paper in Nature Chemical Biology Find the AsCas12f1 plasmids   Selective... Mike Lacy The retroviral-like protein PEG10 specifically binds its own mRNA to facilitate its packaging...of cargo sequences in the capsid, in an approach called Selective Endogenous eNcapsidation for cellular... the prime editor, yielding PEmax, to work synergistically with epegRNAs and PE4 and PE5. These enhanced...systems improve gene editing performance at therapeutically relevant gene targets and expand the application...

...and success. Customer and technical support Our customer and technical service brands continue to ... We score ~99.9% helpful and the customer and technical service teams put 100% effort into helping our...positive, supportive culture, we are attractive to the local employee pool and we get a lot of successful candidates...connections and even via Twitter. We love working with local organizations like Just-a-Start, the Northeastern...

... department at the University of Massachusetts Medical School. We are working with postdocs at other institutions... and all 6,000+ postdocs at the University of California, who had a contract - negotiated by their union... December 1st. Salary inconsistencies across geographical regions When we originally started gathering...salaries. Postdocs at the Salk Institute in La Jolla California are not getting a raise. Meanwhile, in less densely...changes in science. Follow him on Twitter @BiophysicalFrog and Future of Research @FORsymp.   Additional...

...donating and receiving labs)” — Jim Woodgett, biomedical researcher at Lunenfeld-Tanenbaum Research Institute...reagents and protocols so easy!” — Nina Jain, Harvard Medical School. We want to give a special thanks to the...decade. We did this the old-fashioned way: through physical outreach, visiting organizations, knocking on ...standardized agreements and electronic signatures to dramatically speed up MTA approvals. In 2013 we were thrilled...for being Addgene. You guys rock." — Andrew York, Calico Life Sciences. Most of all, we’re incredibly thankful...

...present in the cell, it can also introduce non-physiological RNA-protein interactions due to sample processing...its rate of biotinylation. The improved enzyme, called BASU, is derived from a BirA* homolog found in ... interactions in events that occur on a short timescale, such as during drug action or cell signaling....t be used to study endogenous RNAs at their physiological expression levels; however, knock-in of BoxB... by 90%, suggesting that QKI is a host protein critical to Zika replication. Human genetic disease. RaPID...

...requisite for a functioning lab, but it is for a scaleable lab. When you need to track the location, quality...our second example we’ve added some simple but critical logic. Some steps fail “hard”, they reset the ...interchange them easily as needed. In this case, the physical considerations of the laboratory make object composition...science_name`. You'll need to decide given the technical details of the problem you're solving, but consistency...interesting biotechnology code.There are valid technical reasons to choose a non-Python stack, but if you're...

...sequence gives the loxP site directionality, and the canonical loxP sequence is ATAACTTCGTATA-GCATACAT-TATACGAAGTTAT...the two strands are ligated, resulting in the reciprocal crossover event. This event can have three general... features with loxP sites on either side (often called a “lox-stop-lox” or “LSL” cassette) upstream of...putting the loxP sites on either side of a gene (called “floxing”, for “flanked by loxP”), will permit ...where "N" indicates which bases may vary from the canonical sequence. Among these are loxN (GtATACcT), lox2272...

...and on self-propagation by reproduction. Both physiological processes are controlled by deliberate and compulsory...receptors (NRs), their coregulators, and their physiological endocrine ligands. Nuclear receptors and their...the diet. Additional members of this family are called orphan receptors because their ligands remain unknown...circadian regulation, as well as in important physiological states such as adipogenesis and macrophage activation...widely used today.  The sheer abundance of these critical research tools is a double-edged sword, however...

...methyltransferase DNMT3A allows researchers to easily add biochemical marks to genetic regions from which where they...eventually, develop treatments, we need tools to specifically edit methylation sites.  Step aside, transcriptionally...interact with transcriptional machinery to modify local gene expression, confounding any potential conclusions... DNA. When dCas9 is bound to a target CpG, it physically blocks DNMT1 from that site. So, when DNMT1 methylates...Szyf 2021.   Let me leave you with a few practical considerations: Design your gRNA to overlap the...

... from E.coli, named TadA7.10 in ABE7.10)  a catalytically impaired Cas protein (dCas or Cas nickase)  ... the eighth generation of adenine base editors, called ABE8 (Gaudelli et al., 2020). The generated ABE8s...accidental insertions and deletions (indel) by the catalytically impaired D10A nickase from ABE7.10 can be a ...the authors created ABE8 constructs using a catalytically “dead” S. pyogenes Cas9 (dC9-ABE8.x-m/d). The...even in adult patients. In order to analyze the clinical use of ABE8 for primary human cell editing, the...

... staying on top of the ever expanding body of biomedical literature! PubMed PubMed is the classic search...interest.  Best uses Accessing the vast body of biomedical research papers. Twitter Scientists often use...papers, and more. Saving papers There isn’t technically a way to save papers on Twitter, but you can ... contain a short summary of the topic and a chronological list of highly cited papers that contain your...also just follow feeds curated by Meta. Feeds automatically update as papers are published, similar to Facebook...

...expression observed with wild type CoChR, soCoChR localizes primarily to the cell body with very little signal...engineered microbes, used coffee grounds could be economically repurposed as starting material for the bioproduction... FLINC biosensors for live-cell imaging of biochemical activity Article contributed by Angela Holmes... live-cell imaging of the dynamic changes in biochemical activity within the cell. Their Fluorescence ...peak of 519 nm. These proteins have improved photophysical properties and can be used in quenching, FRET...

...past decade, primary cells remain difficult to genetically edit due to toxicity from electroporation or ...some cases) and minimal toxicity, including in clinically-relevant lines such as CAR-T cells. Plus, this...Cas9, fused to HIV TAT, Myc and SV40 Nuclear Localization Signals, and GFP. B) Workflow to assess Cas9...microscopy in HeLa cells expressing COX8a-mStayGold. Scale bars 10 μm (main), 1 μm (inset). Image reused from...versions. Thanks to Christophe Leterrier, whose FocalPlanes piece on mStayGold inspired and informed this...

...the transfection method (i.e. polyethyleneimine, calcium phosphate or liposomal methods), the volume of ...unconcentrated prep. For example, at Addgene we typically concentrate 100-fold meaning that a 10 cm2 dish...experiment requires a certain volume of virus, consider scaling up by either increasing the number of 10 cm2 dishes...be optimized prior to starting. We recommend calculating the ratio of the surface area of the larger vessel...lentivirus is the cell line that is used for titering. Typically researchers will choose a relatively permissive...

...can contain different neurotransmitters. Using genetically encoded fluorescent proteins coupled with Fibronectin... endogenous synaptic proteins allowing them to localize to the synapse and minimize extraneous fluorescence...Jerome Goudeau, Cynthia Kenyon, and colleagues at Calico Life Sciences. Like many split FPs, split-wrmScarlet... genes whose protein products have distinct localizations. Figure 2: Split-wrmScarlet labeling ...AAV5. Find the AAV prep here The red-shifted calcium sensor jRGECO1a is now available as AAV-PHPeb. ...

...using either loxP or lox511 sites, leaving two identical sites on one end of the DNA fragment (Figure 1...recombination event then excises the DNA between the identical loxP or lox511 sites (Figure 1, bottom). Since...presence of Cre recombinase. This plasmid now specifically drives expression of mCherry instead of BFP.... rescue with in the antisense orientation (let’s call the gene “GeneR”). The key to this experiment is...information. References: 1. Schnütgen F, Doerflinger N, Calléja C, Wendling O, Chambon P, and Ghyselinck NB. A ...

...workplace, and pursue leadership opportunities at the local, institutional, regional, and national levels. These... first at Brigham and Women’s Hospital/Harvard Medical School and then Boston University (institutions...issues faced by postdocs and institutions are not local but are national issues. I applied and was successfully...other people mostly online and on monthly phone calls. This has its challenges as meeting in person is...Chair then stepped down, I became interim Chair automatically as it was an understood expectation of any Vice-Chair...

...and solutions to research problems. This movement calls itself the Open Science movement and is part of ...and the reagents are too expensive to make it practical - but we have members looking at how to re-use...don’t throw old equipment away until you ask your local makerspace if they want to hack new life into it...somewhere near you. The place to start looking for your local space is the Hackerspace website. Markerspaces differ...fun with technology, so feel free to contact your local space and get involved! Derek Jacoby is a PhD ...