...to learn much more about cellular DNA repair pathways. An emphasis on cellular repair post-CRISPR cleavage... a common thread at the meeting. The new Somatic Cell Genome Editing program, run through the NIH Common..., which alter genomic DNA without the need for cellular HDR.   Having had time to digest my notes, since...Improve HDR Efficiency for CRISPR Editing in Human Cells Hands-on tips for repair template design from guest...

...these issues by creating immortal antibody-producing cell lines (hybridomas) that secrete antibodies in culture...reproducible production. However, even clonal hybridoma cell lines can produce more than one mAb (Bradbury et...about recombinant antibodies in this blog post. Hello, NABOR! Addgene will bring the same open-access ...resource of plasmids encoding recombinant antibodies as well as other affinity reagents such as single-chain ...need for an open-access, molecularly defined, and well validated collection. I’m thrilled to be working...

...R-factors could be transferred between bacteria via cell-to-cell contact; however, scientists noted that, unlike...cytoplasmic factors that could be transferred between cells. At the time, these extranuclear agents of heredity...PMCID: PMC3785787. References: Lederberg, Joshua. "Cell Genetics and Hereditary Symbiosis" Physiological...Transmissible Drug Resistance in an Epidemic Strain of Salmonella Typhimurium.” The Journal of Hygiene 60.3 (1962...

...to supplement your cells with biliverdin for maximal brightness. In mammalian cells, one of the improved...Imaging Center at UCSF. A common requirement for live cell imaging experiments is the ability to follow multiple...emission spectra of the protein, so that the protein is well excited by your microscope and the fluorescence ...laboratory of Ronald Vale, after which he was a fellow at the Bauer Center for Genomics Research at Harvard...

...team potential. I often thought that performing a cell culture experiment myself would take less time than...Another team member was highly self-motivated and excelled when given the freedom to explore projects autonomously...management skills. Finally, developing emotional intelligence is vital. Learning to "read the room," understanding...part of your broader career development.   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...

...recognized and valued for its immense importance, it may well be what determines whether you get the job after...as an opportunity to practice communicating.  Excellent scientists learn from advisers and other mentors...forget that you have a much bigger job than merely telling your audience about the work you've done. If you...help graduate students in all STEM fields become excellent science communicators. To empower graduate students...in science communication The strength of story telling Science communication: 9 strategies to get your...

... Many thanks to our guest blogger David Mellor! David Mellor leads the policy and incentive programs ...This post was contributed by David Mellor from the Center for Open Science. In the last decade, researchers...Before coming to the Center for Open Science, Dr. Mellor worked with citizen scientists to design and implement...at https://orcid.org/0000-0002-3125-5888 and @EvoMellor.  ...

...can also find Dr. Page and Hello PhD on Twitter: @ThePurplePage and @HelloPhD "Role" call. #BLACKandSTEM...Science Communication” at the American Society for Cell Biology (ASCB) 2018 Conference. I don’t exactly ...Scientists SHOULD Be Using Twitter" podcast from Hello PhD In this podcast the hosts, Josh and Dan, discuss...there’s some great reminders for Twitter pros as well, like tip #4: “Follow people for about a week and...

...have lunch or coffee with someone I don’t know very well at least twice a week. I have done this with people... career advice. Your family and friends know you well and might have good ideas for who you should connect...Healthcare Businesswomen’s Association hosts an excellent group mentoring program in many of its chapters...keep working with you is report back with progress. Tell them how their advice made a difference, helped ...in a leap in your skills. If a first meeting goes well, you can say, “This was really helpful, perhaps ...

...thinking. Being able to learn things quickly. Story telling. The ability to know what’s a story and how to ...to hook your readers with it.” She feels that storytelling is an impactful tool for science communicators...through connections and that’s something that good storytellers in science can do.” Susan’s number one piece... starting a blog, but with one caveat: “I hate telling people to work for free because you shouldn’t work...social media as a scientist here Learn more about storytelling in science ...

...system, antibodies are produced by B cells. Each individual B cell produces antibodies that all recognize...also all the same isotype. But together, all the B cell clones in the immune system make different isotypes...antibodies starts with selecting just one of these B cell clones for further antibody production, polyclonal... blood from the immunized animal. The red blood cells and the serum are separated, and the serum - which...

...Professional Career Coaching can be an excellent tactic for scientists making a career shift or who wish...of this to get the information you need to choose well. What are their qualifications? Do they have certifications...does not endorse any of these coaches and I can't tell you which one will be a match for your needs, but...Consulting is part of that group and she is especially excellent in coaching for communication skills. Industry...Industry transition is also an area of specialty for Michelle Capes at Adeptify Me. While based in Madison, ...

...encode proteins. They play important roles in many cellular and regulatory processes, but much of their functionality...RNA (sgRNA) to create a double-strand break works well because very small deletions and insertions in the... should be able to devise an experiment that can tell you more about its function.  Resources and references...screening for functional long noncoding RNAs in human cells by Cas9 targeting of splice sites. Nat Biotechnol...non-coding RNAs with the CRISPR/Cas9 system in human cell lines. Nucleic Acids Research. 2014;43(3):e17. doi...

...better than a nitrocellulose membrane. If you have already run your blot using nitrocellulose, you may decide...protein of interest in a complicated solution like cell lysate. But they can be a lot of work — and what...instructions for the membrane you are using. Dry nitrocellulose membranes are brittle, so proceed with caution...to solutions. It is also volatile and toxic (and smells bad!), so be sure to use it under a fume hood. ...

...plethora of antibody options for your experiment (all well validated for your application, of course!) When...the isotype, and subclass affect my experiment?” Well, it depends on the application and target. If you... blot and probing a single target that expresses well, then probably not. In this case, take a look at...degree of affinity, then species choice could very well matter. Take rabbit versus mouse monoclonal antibodies...for Light and Electron Microscopy. In Methods in Cell Biology (pp. 283–310). Elsevier. https://doi.org...

... question about what should go into a cell, hover over the cell and instructions will pop up. When you...spreadsheet (Figure 2) and are described in Table 1. Some cells have drop-down options for you to choose from, while... into the appropriate full and partial sequence cells. *Note: If you believe you need to deposit a collection...Relevant Mutations Indicate the amino acid change as well as any functional consequences of that change. For...

...currently doing this using two approaches: single-cell RNA-sequencing to identify neuronal class and deep...Drosophila and zebrafish articles published there as well.   Worm Development Dynamics Database (WDDD) The...WDDD aims to acquire quantitative data on all the cell divisions of the C. elegans embryo when all essential...Wang et al., 2018).” With the split system, only cells that have both halves of cGAL can drive expression... Alyssa Cecchetelli...

...since the lab was highly focused on protecting intellectual property, it worked away in isolation. It published...joint. Silly ideas (mining local salt deposits to sell at farmers markets), and serious ideas (how to stop...color-changing flowers was a great business idea as well, and they thought we could be taught. They invested...Biochemistry Department at the University of Manchester as well as the Biology Department at CSU. When not pursuing...enjoys spending time with her husband and two Jack Russell Terriers. More SynBio Reading Read more Addgene...

... for Mapping Multiprotein Complexes in Human Cells.” Cell Rep. 13: 621-633. PMID: 26456817. Find plasmids...Chromosomes). Using their PITCh plasmids, GFP knock-in cell lines can be created in about a month and a half...locus-specific gRNAs Step 4: Select for puromycin resistant cells Step 5: PCR amplify and sequence the locus to verify...crispr.mit.edu. They tested their system in HEK293 cells, integrating the GFP-Puro cassette into the FBL ...

...overexpress YFG in a Drosophila cell line. Knock downs in Drosophila cell lines are extremely effective....days after setting up the genetic cross). You can tell when pupae are ready to hatch the next day because...person. Animals are diploid except for their germ cells (sets of genomic material are contributed by mother...various fly reagents on the Bloomington website as well. Additionally, they have useful information on topics...

... in mammalian cells Hu et al. compared xCas9 activity with wt SpCas9 in mammalian cells, finding xCas9...ABE7.10, as well as improvements at NG and GAT PAMs. xCas9 3.7 Activity in Mammalian Cells Construct... percent editing efficiencies (edit frequency in cell population measured with NGS) are shown as percentages...

...doing a brief academic postdoc. Why did I switch? Well, my long-term goal was to enter industry since I...” that you’ve sold out. You’ve given up your intellectual freedom and integrity. (Not the case, as I’ve...s ability to manage others and work as a team as well as their scientific abilities. At AstraZeneca, scientists...who get postdocs and do not support or manage them well are simply no longer allowed to have postdocs. In...Scientist at AstraZeneca. She has a background in cell and developmental biology and is applying this towards...

...encoded tools, such as microbial opsins, to control cellular activities using light. In 2015, scientists combined...activator domain (VP64 or p65). Upon expression in the cell, the dCas9-CIB1 fusion binds to the target DNA sequence...activator domain VP64. Using this system in HEK293T cells to induce expression of human IL1RN, the researchers...protein with three red fluorescent tags in U2OS cells. Upon light activation, dCas9 is released within... silencing of transferrin receptor CD71 in HeLa cells.   Comparison of photoactivatable CRISPR strategies...Spatiotemporal Control of CRISPR/Cas9 Function in Cells and Zebrafish using Light‐Activated Guide RNA. Angew...

...Nanobodies are like tiny antibodies which work just as well, if not better, than antibodies for all of the above...structure and function compare to IgG antibodies, as well as how to produce them for use in your lab. Structural...triple colocalization is possible even when staining cells with three IgG1 antibodies. Antibodies were individually...workflow yields similar localization patterns as cells stained for one target (compare nanobodies staining...–rabbit IgG secondary nanobodies. The Journal of Cell Biology. PubMed PMID: 29263082 Find plasmids from...

...machinery, and could thus be used in non-dividing cells. Cas13 enzymes also don’t require a PAM sequence... RNA knockdown. In initial tests, Cas13b from Prevotella sep. P5-125 (PspCas13b) yielded 62.9% average... a maximum of 28% editing efficiency in HEK293T cells, as assessed using RNA-seq. Since the REPAIRv1 machinery...including reversibility and use in non-dividing cells. With the expanded targeting ability using both ...Functionally Orthogonal Type VI-A CRISPR-Cas Enzymes." Mol Cell. 66(3) (2017):373-83. PubMed PMID: 28475872. Find...

...not generally in a cell’s best interest to overexpress a protein. Energy and cellular resources are being...being spent to make something the cell doesn’t need to make. Eukaryotes and some bacteria deploy proteosomes...proteosomes to degrade what the cell might consider junk protein. Though there are a number of chemical and...same manner as it would have in the context of a cell. FLAG tags can be cleaved by enterokinase, which...

...day care and school if you tell them they will enjoy it. If you cry and tell them they will miss you, of...having anyway) for enjoying your career and life as well as your family and friends. If there is any guilt... way more fun. And despite those who may wish to tell you so – towel corners do not need to match up exactly...example, say no to some of those birthday parties. Tell your mom you can’t make it to dinner this week. ...

... 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...

...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...

...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...