...four proteins Oct4, Sox2, Klf4, and cMyc (the so called OSKM factors). As the Yamanaka lab's primary Addgene...field has exploded so quickly, citing the first clinical trial with iPSCs with his collaborator Takahashi...picture perspective: Dr. Yamanaka talked about his clinical trial patients with advanced macular degeneration...Eggan presented work that is leading to promising clinical trials for ALS.   I have an uncle with Parkinson's...the first stem cell line that could be used in clinical trials. The quality control process alone takes...

...made of chemicals, which have an unfortunate tendency to react with other chemicals and biological structures...antibody out on the bench overnight. Do not panic! Typically, antibodies are okay if they’re left out overnight... fluorophores, which work not by reacting to a chemical, but by emitting a specific wavelength of light...

...Bryan Roth’s lab created an open source resource called PRESTO-TANGO (parallel receptorome expression and...activated downstream of GPCRs. This collection, called TRUPATH, allows researchers to identify specific...neurons and specific behaviors. Chemogenetics uses genetically engineered receptors to interact with small molecules... to optogenetics but instead of light, these genetically modified receptors are activated by small molecules...some GPCRs further to respond to specific, pharmacologically-inert, small molecules. These new receptors...

...professors, and mentors. Ask for specific guidance on grammatical errors, sentence structures, and overall format...support.        5. Application submission  Set a calendar reminder for every deadline and shoot to submit...Pierre is currently a doctoral candidate in the Chemical Biology Program at Harvard. Roo wants to simplify...simplify access to academic resources among economically disadvantaged populations. Roo is the co-founder...resources that can help their children thrive academically. Rose is the co-founder of STEAMid.     Additional...

...plasmids can be used to carry out any biochemical and biophysical study involving protein stability or ...degradation. Despite how easy these experiments are to physically set up in the lab, RNAi is often completely ineffective... most scientists use mutant animals that are genetically altered to overexpress endogenous RNAi pathways...facilitate protein-protein interactions and mediate biochemical processes. One of the largest and best characterized...

...nearly 50%, showing proof-of-concept for pre-clinical and clinical applications of dCasRx. The identification... with CasRx and AdmCas13d fused to a nuclear localization signal (NLS). In a HEK293 mCherry reporter assay...target essentially any RNA in a cell. Since catalytically dead dCasRx maintains its RNA-binding properties...enhancers. After AAV-mediated delivery to iPS-derived cortical neurons, dCasRx-mediated exon skipping improved...

...robust, with an eNL-based calcium sensor displaying a 500% signal change upon calcium binding. Suzuki et al...around these issues, but this solution wasn’t practical due to the low intensity of luminescent proteins... to previous publications. They also developed calcium, cAMP, and ATP sensors based on Nano-lantern, showing...not alter cellular physiology in your system. Optical sectioning cannot be conducted with luminescence...

...plasmid with the delivered cargo (often a gene) is called the transfer plasmid while the other plasmids with... with packaging or replication components are called helper plasmids. Below, we’ll breakdown the vector...viral experiments is to deliver a genetic cargo, typically an expression construct, to cells/organisms. This...yield dsDNA. Unlike LTRs, ITRs are perfectly symmetrical to allow for this pairing to occur. Like LTRs...gene, making it replication deficient. This is typically supplied by the production cell line (HEK-293)...

...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...