...off-target effects. The flaws of RNAi screens are well characterized and strategies exist to control for...delivered as a pooled library via a lentiviral vector to cell lines that either stably expressed Cas9 or that ...Hart et al, Bassik et al). The library performed well in both screens, with the growth screen having a...almost all genes involved in the production of a cell surface glycan that’s required for ricin uptake....reveals pathways underlying ricin susceptibility." Cell 152.4 (2013): 909-922. Pubmed PMID: 23394947. Pubmed...

...particles that can transduce cells. Infectious titers are typically quantified by cell transduction assays. Wild-type... on three important factors. What can AAV titers tell you? Physical titer: the concentration of viral...arrowhead). Scale bar = 100nm. Image courtesy of David Bell and Svetla Stoilova-McPhie, Center for Nanoscale...Briggs HA, Gallichotte EN, Ruf IK, Hindson BJ, Vessella RL, Tewari M. Absolute quantification by droplet...

...Specifically, roles in technical support make excellent use of the skills developed by life science PhDs...issue was with a custom-made reagent to manipulate cells (manufactured by one of the larger biotech companies...patient enough to find the real issue, you will do well in a technical support role. What general characteristics...go wrong in a lengthy protocol. And customers may tell all sorts of details about the application, just...about novel developments in the fields of gene and cellular therapies.     Additional Resources on the Addgene...

...how do proteins X and Y regulate the Z pathway?" A well-defined outcome makes it easier for the search committee...example, "while it is possible to grow liver stem cells and liver-like mini-organs or organoids, neither...combine bioengineered scaffolds with patient stem cells to generate compatible functional livers.” Why are...Health Sciences University, did his postdoctoral fellowship with Jennifer Lippincott-Schwartz at the National... optimization of fluorescent proteins,  and live cell imaging approaches. Erik is also an avid long distance...

... grammar and spelling several times and ask someone else read your cover letter (as well as your resume...“I’d be good at that! I organized my lab data so well that I can find the results of any experiment I’...of experience in [field of research], and have excellent [list the most important skills for the position... specific examples. Instead of saying “I have excellent communication skills” try giving an example, like...

...convenient and effective, in both academic circles as well as in industry. The news is good. In their ideal...their data in one place and avoid scattering intellectual property across various software providers used...:   Does your lab need to keep paper records as well as electronic ones? It is important to know in which... it is because you need to keep paper records as well as digital, or for reporting on a project – printing... or company, site licenses might be available as well. In most cases this applies to universities. How...

...because of its gain in popularity. Gibson assembly is well known for allowing easy assembly of multiple linear...is immediately ready to transform into competent cells. The master mix of enzymes can be purchased from...mix for 1 hour, and then transform into competent cells. The success rate of this reaction is typically ...polymerization steps could occur. Secondly, it does not work well if the ends of the fragments have stable single ...and ordered, and are specific for each fragment as well the fragment you want next to it, so this doesn’...

...directly by creating well characterised antibodies against all human extracellular proteins with community-based...are quite low (<30%, Prinz et al 2011, Begley and Ellis 2012). Recent efforts from the Reproducibility Project...Hidden “negative” data We all want to be able to tell a good story when presenting scientific findings... results. Not only is it scientifically and intellectually affirming to have a hypothesis confirmed, it...

...annealing to a complementary ssDNA target in the cell.  Only Beta expression is required for recombineering...fragments, such as antibiotic resistance genes, as well as non-selectable DNA fragments, such as gene replacements...definitely the easier of the two options, but these cells are prone to mutations and will have more unintended...Central PMCID: PMC307639. 2. Daiguan Yu, Hilary M. Ellis, E-Chiang Lee, Nancy A. Jenkins, Neal G. Copeland.../Knockin Plasmids to Disrupt Genes in Mammalian Cells CRISPR Methods for Bacterial Genome Engineering ...

...inspired. CAUTION: Spoilers alert! Pre-DNA Era  H.G. Wells, the father of scientific fiction, was also a trained...Asimov, Arthur C. Clarke, James Blish, and John W. Campbell. This period is termed as the golden age of SciFi... in the genome. These technologies rely on the cellular DNA repair machinery, which can lead to high frequency...1258096 . https://doi.org/10.1126/science.1258096 Gaudelli NM, Komor AC, Rees HA, Packer MS, Badran AH, Bryson... C (2000) Hugo De Vries: from the theory of intracellular pangenesis to the rediscovery of Mendel. Comptes...

...temperature incompatible to genome editing in human cells. The Feng Zhang lab identified a Cas12b from Bacillus...can block Cas12a-mediated genome editing in human cells. Read the paper in Science Find the anti-CRISPR...anti-CRISPR expression plasmids CRISPR-based method for cell lineage tracing Valcyrie is a CRISPR-based marker...marker excision method for marking distinct cell clones developed by the Michalis Averof lab. It’s an alternative...

... (iPS) Cells - 2008 was marked by the influx of plasmids for reprogramming differentiated cells into pluripotent... pluripotent stem cells. Plasmids from Shinya Yamanaka's lab and James Thomson's lab were the most frequently...infographic created by our own Addgenie, Laura Veckerelli. Click on the picture below to see the full graphic...

...Clearly we have all of our writers to thank for the excellent content they’ve churned out in the last month....contributed by guest blogger Kristian Laursen from Cornell University, is a bit old school and provides a ...or G->A transitions in a target sequence in live cells. This technique uses catalytically inactive CRISPR...and other means of generating mutations in live cells are all discussed further in this informative post...

...down to one, key concept: Creating a culture of excellent, effective communication between all members of...ensuring that the group always conducts exceptionally well-managed meetings. Members face one another, and...engagement in the issues at hand and eagerness to get, as well as give, information to make improvements. Members...meetings with team members (don’t become known for cancelling these); ask for feedback and input on multiple...

... the News Chapter 1: Introduction - Vini and Amy tell us a little about themselves. Chapter 2: What is...brings in scientists from a variety of fields as well as students from the ed school, design school, and...context. Episodes are driven by narrative and story telling. Chapter 5: The SITN Blog - Vini and Amy describe...scientific information for the general public as well as a platform for scientists to be viewed as people...

...goals/needs/situation in mind, rather than (or as well as) their own.” “Someone to act as your ‘cheerleader...Apparently, a good Jedi does not necessarily an excellent mentor make. Are your current mentors/advisors...concrete goals and helps you reach your goals Listens well and communicates clearly and openly Gives honest...What Kind of Mentor Do I Want To Be?” In this excellent essay Acclimatrix, an assistant professor at a...

...make-over as well. While in the course of redesigning the homepage, and striving to tell our story in ...navigation, while also updating brand messaging, as well as simplifying the page, and making it all look ...content to dictate the space. We wanted to comfortably tell our story, to get scientists excited about what ...

...we'd like to extend a special thank you to Dr. Tom Ellis (Imperial College London), Dr. Connie Cepko (Harvard...asked leading professors for their top 3 tips. Tom Ellis 1. Keep up with the literature. He recommends setting...mentees who have goals outside of the university? Ellis recommends that a student interested in leaving ...Acknowledgments Thank you again to Connie Cepko, Tom Ellis, and George Church for answering our burning questions...

... GFP, enabling you to select cells with high GFP expression. These cells have the highest levels of Cas9... on the same plasmid, you’ll make sure that each cell that gets the plasmid contains all of the desired...gRNA, which causes hypopigmentation, thus marking cells that have undergone genomic modification. In this..., et al. “Multiplex genome engineering in human cells using all-in-one CRISPR/Cas9 vector system.” Scientific...

...Signal/Imaging: Fluorescent reporters are not as well suited for imaging organs located deep in the body...long vesicles have hollow interiors and a protein shell that excludes water but is gas permeable. These ...These vesicles are primarily used by water-dwelling photosynthetic bacteria and archaea to regulate their ...limit of detection for arg1 expressing ECN is 109 cells/ml, which is within the range of  the ~1011 commensal...

...human body, cellular biology, medical specialities, and general items. Individual images as well as collections...website has many tutorials and Lynda has a tutorial as well, which you may have access to through your university...discussed? Or know of a tool or resource not mentioned? Tell us about it in the comments section!  Additional...

...antigens in living cells or tissues. sdAbs can also be used to manipulate cellular functions, like in ...sdAbs over conventional antibodies include:  Excellent tissue penetration, thanks to their small size... their way out of the lab and into the clinic as well. The first sdAb based therapeutic was approved in...

...tangential subjects, and let curiosity guide your intellectual journey. Practical approaches One approach...outdoors in nature or visiting museums can be excellent methods to detach from the constant stream of ...unlocking the next frontier of knowledge. Matteo Tardelli authored the book "Beyond Academia: Stories and...molecular biology and metabolism. He now guides fellow PhDs in their careers with actionable steps and...

... repository of CRISPR screens Cellosaurus: A knowledge resource for cell lines IBEX Knowledge Base: A ...publications and commercially available sources, as well as those deposited with Addgene. NCBI BLAST: Compares... stand-alone sites for general primer design, as well as primer design for different cloning applications...

...50% of predicted genes have unidentified roles as well (Vanni, et al). Is it true that this many genes ...fluorescent protein (GFP) was the reporter gene and green cells were observed in the kidneys at a specific developmental...genes of interest may not be expressed in the tissue/cell line you are working with. Development-specific...10.7554/eLife.67667 Springer, S. P. Gene Traps. Plant Cell. (2000) 12(7): 1007-1020. DOI: 10.1105/tpc.12.7.1007...

...scientists to get out there and talk about their work. To tell others why that work is cool and why it really matters...seem all that interested in the details, work on telling it another way. You’ll know you are on the right...Plan to write regularly and, when you are ready, tell your friends or others in the field where to find... science writing, look into a AAAS Mass Media Fellowship or maybe an internship at a place like Science...

...devise new strategies to reproduce within a host cell. A few virus particles have learned to circumvent...deliver a highly specific, programmable platform well-suited for various innovative biological and translative...2011 Jun; 9, 467-477. Bondy-Denomy J, Pawluk A, Maxwell KL, Davidson AR.  Nature. 2013 Jan; 493, 429–432...): 819–23. Mali P, Yang L, Esvelt KM, Aach J, Guell M, DiCarlo JE, Norville JE, Church GM. Science. ...

...Nanobody toolkit to monitor retrograde transport at the cell surface Article contributed by Marcy Patrick  Listen...nanobodies to study retrograde transport from the cell surface. Their toolkit includes a set of plasmids...new technologies to quantify proteins in tissues, cells, and biofluids. Recently, Sam Myers, a Scientist...an sgRNA expressed from a separate plasmid. When cells are treated with biotin–phenol and H2O2, APEX2 labels...

...closely with TTOs to help protect our depositors’ intellectual property, and to help requesting institutions... research; provide specific examples if possible Tell them about the Golden Stopwatch! Any organization...University  Princeton University  Purdue University Rockefeller University  Salk Institute for Biological Studies...agreement Technology Transfer: the exchange of intellectual property (as inventions, ideas, plasmids) among...

...thermoplastic polymers that can work with all the bells and whistles. The fibers are made from a process...later! Next, additional components are added - the ‘bells and whistles’ – including microscale light emitting...of the stiffness the fibers would need to be, as well as other features necessary for implantation in ...and chemically excited a class of enteroendocrine cells within the intestines known as neuropods. They observed...