... that many researchers were having difficulty achieving indel formation (gene disruption) with this new...most of the respondents have been struggling to achieve genome editing with NgAgo, this does not mean the...successful indel generation and epitope knockin was achieved by a few respondents suggesting that NgAgo can...most of the researchers surveyed here could not achieve indels with the NgAgo system indicates that it ...which might be rapidly degraded by the cellular machinery, leaving a small amount of active NgAgo/ssDNA ... cell. As proposed in the original article, to achieve greater efficacy, researchers will have to carry...

...for in vivo delivery of advanced genome editing machinery such as CRISPR-Cas. Figure 1: Schematic...transduced to target cells. Within target cells, Cas9 machinery is transiently expressed and binds to target DNA...rates of targeted mutagenesis similar to those achieved by isogenic AdVs encoding TALENs targeting the...). If sufficient gene transfer and knock-in is achieved, producer cells could produce corrective amounts...potential platform that could be realized via simple switching of the relevant gRNAs and donor DNA Adenoviral...Han Y-C, Ogrodowski P, Crippa A, Rekhtman N, de Stanchina E, Lowe SW, Ventura A (2014) In vivo engineering...

...domain. Feng Zhang's group utilized the bi-lobed architecture of Cas9 to engineer a series of "split" cas9...must not act on any of the other bits in order to achieve accurate and reliable multi-bit memory. To this...cassette, you can specifically express your new chimeric protein in a variety of model systems. The MultiSite...but a translated mRNA will be red. This system achieves great specificity and resolution by using 6-12...mitochondrial subpopulations within the same cell; by photoswitching PSmOrange and photoactivating PAGFP in separate...screened for speed and voltage sensitivity. This hierarchical screen led to the generation of two enhanced... CMV-LUC2CP/intron/ARE). The Luc-I intron is a chimeric β-globin/immunoglobulin intron that has been commonly...

...become popular. There are only so many methods and machines one team can learn so linking together expert ... a mailing list. This project gave me practice reaching out to other scientists, being clear about what...beyond your organization and/or scientific field. Reaching out to people who have little in common with you...starting a collaboration with a colleague. However, reaching outside your knowledge sphere allows you to connect...time to talk or to at least read my emails. Then pitching myself and the event was very difficult and somewhat...experience during my graduate studies in regard to reaching out to new people, checking in regularly, and ...

... plasmids to utilize diverse host replication machinery. For instance, the ori typically contains iterons...However, if plasmids have their own replication machinery such as Rep or other initiator proteins, they ...vulgaris, Erwinia carotovora, E. chrysanthemi, Escherichia coli, Gluconacetobacter xylinus, Gluconobacter...E. herbicola, E. rubrifaciens, E. stewartii, Escherichia coli, Klebsiella spp., Legionella pneumophila...spp., Brucella spp., Caulobacter crescentus, Escherichia coli, Gluconacetobacter xylinus, Paracoccous ...integron promoter-based expression systems for Escherichia coli and Agrobacterium tumefaciens. 3 Biotech...

... passes through the machine’s lasers and gives off a signal read by the machine’s sensors. (Usually this...or unbound antibodies.) Each event read by the machine is plotted as a dot on an X,Y axis to generate ...variations in fluorescent signals, and how the machine reads/interprets the signals, the axis is instead...populations. This shape is called a ‘gate’; it tells the machine that everything inside the gate is one population...Doloff, J., Bose, S., Vegas, A., Ma, M., Sahay, G., Chiu, A., Bader, A., Langan, E., Siebert, S., Li, J.,...

...Sommella A, Colecchi L, Rossi S, Simonelli F, Giunti M, Bacci ML, Polishchuk RS, Auricchio A (2015) Improved...genes into AAV First, some history. In 2008, the Auricchio lab decided to solve the issue of “tiny vector...tested and with the optimal sequences, the group achieved ~80% of muscle cells expressing mini-dystrophin...Cutillo L, Sparrow JR, Williams DS, Bennett J, Auricchio A (2008) Serotype-dependent packaging of large..., Giunti M, Palfi A, Farrar GJ, Polishchuk R, Auricchio A (2013) Effective delivery of large genes to ...

...typical PCR machines, the authors use affordable, field-ready, Bluetooth-enabled PCR machines that can even...approximately 1-2 hours Equipment needed: Mini PCR machines by DIY-Bio, P51 cardboard fluorescence visualizer...mentioned, likely ~1-2 hours Equipment needed: PCR machines/Heat blocks, Lateral flow strips, Agarose gel .../2020.04.07.028167 Broughton JP, Deng X, Yu G, Fasching CL, Servellita V, Singh J, Miao X, Streithorst...Miller S, Pan C-Y, Guevara H, Wadford DA, Chen JS, Chiu CY (2020) CRISPR–Cas12-based detection of SARS-CoV...

...tag. Many thanks to our guest blogger, Joachim Goedhart! Joachim Goedhart is an assistant professor at ...This post was contributed by guest blogger Joachim Goedart, an assistant professor at the Section of ...living cells and organisms. Both components of the chimera are encoded by DNA. Since researchers can generate...from a plasmid-based system to assess whether the chimera is still functional. Final words on fluorescent...fluorescent probes. You can follow him on twitter: @joachimgoedhart.   References Adjobo-Hermans, Merel JW, et ...

...scientists and non-scientists alike are marching for. I will be marching to advocate for science education ...is a Pro-Science and Pro-Evidence march. Who is marching and why? Who? All sorts of people are choosing...promises more cuts (3, 12, 13, 27). In order to achieve the breakthroughs, technologies, and treatments...exhaustive list and every marcher is not necessarily marching for every cause. The references provided also ...opinions or research on each topic. Resources for marching and advocacy If the march for science rallies ...

...Addgene Board of Directors. Cicalese served as the Chief Human Resource Officer for Harvard Medical School...Giving, an organization that raises money for childhood cancer through the sale of art. About two years...about Julie Cicalese  Ms. Cicalese served as the Chief Human Resources Officer for Harvard Medical School...Giving, a 501c3 public charity raising funds for childhood cancer through the sale of art. Past volunteer...Marathon for the MGH Pediatric Oncology and the Chicago Marathon to benefit Neurofibromatosis, New England...

...   Are you an only child and used to a lot of attention? Or are you one of 8 children and used to getting... with a new PI. I realized that as a first born child, I would thrive on this level of attention and interaction...role model Make sure the mentors you choose are teaching the stuff you want to know and display character... a mentor that sees you only as a paper making machine. Choose one that is a true mentor, supporting career...collaboration? If you are considering a career at a small, teaching college you might want to consider less expensive...

...understanding the brain’s function is knowing its architecture, in particular the connections between different...two pathways also use different cytoskeletal machineries to facilitate transport: retrograde transport...interaction with uptake or intracellular transport machinery. Addgene offers viral preparations of AAV2-retro...vectors. Improved neuronal tracing could also be achieved by using optogenetically controlled transgenes...neuromuscular diseases it would be desirable to achieve transgene delivery by targeting peripheral structures.... Beier, K.T., Saunders, A., Oldenburg, I.A., Miyamichi, K., Akhtar, N., Luo, L., Whelan, S.P., Sabatini...Kuleshova, E., Ojala, D., Huang, C.C., Gerfen, C.R., Schiller, J., Dudman, J.T., Hantman, A.W., Looger, L.L....

...be needed to achieve your intended next career stage and create a timeline for achieving these goals. ...the position, you may also want some evidence of teaching experience. How do you get there and more practically...not always available for mentees due to travel, teaching, grant writing, etc. Also, it is unlikely that...management and conflict resolution workshops. Get teaching experience and learn about modern pedagogy techniques...can help boost your ego in between scientific achievements. I've found cooking, gardening, hiking, and ...

...Below, you will find some easy talking points for broaching this subject with your institution’s tech transfer...resources (summaries, electronic MTA approval, archive of old MTAs, and more) Does your TTO have a question...of Medicine  Baylor College of Medicine Boston Children's Hospital  Brandeis University  Broad Institute...Laboratory  Dana-Farber Cancer Institute  Fred Hutchinson Cancer Research Center Memorial Sloan-Kettering...Studies The Scripps Research Institute St. Jude Children's Research Hospital  Stony Brook University  University...

...Travel Awards: Melissa Chiasson, Elizabeth Unger, and Michal Bernach. Melissa Chiasson: Quantifying the abundance...time is time consuming and unscalable.  Melissa Chiasson, a graduate student in Douglas Fowler's lab at...from each bin are then extracted and sequenced. Chiasson is currently using VAMP-seq to study vitamin K...scales and making personalized medicine a reality,” Chiasson explains. Elizabeth Unger: Developing a serotonin...

...Founded as a nonprofit in 2011, BioBuilder is the brainchild of Dr. Natalie Kuldell from MIT’s Department .... Built primarily from her 20 years of college teaching experience, Kuldell sought to create a program... begins with students reading comic strips and watching animations that present a specific research problem...test their own project ideas using synthetic biology.Launching with a kickoff meeting in October, BioBuilderClub...innovators that opened in August 2017.  This flexible teaching space offers STEM programming, workshops, and ...

... starting with an established method called ChIP-seq. ChIP-seq relies on antibodies to recognize and bind...been doing that for years using this assay called ChIP-seq, but it’s really hard for lots of transcription... antibody to any protein to get really beautiful ChIP-Seq results. The FLAG epitope is quite small—20 ...done comparisons of our new system versus standard ChIP-seq for transcription factors known to have good...

...conversions are base transversions: they require switching between a two-ring purine and one-ring pyrimidine...experimented with adding different proteins to the CGBE architecture. They conducted a CRISPRi screen to identify...were included in the CGBE. They also developed a machine learning model capable of predicting editing outcomes...gRNA/sgRNA Guide RNA RNA used by Cas9 to locate a matching DNA sequence.   Target strand The DNA strand... CRISPRi screens, target-library analysis, and machine learning. Nature Biotechnology, 39(11), 1414–1425...with sequence context preference predictable by machine learning methods. Nature Communications, 12(1),...

...Many thanks to our Guest Blogger Londa Schiebinger. Londa Schiebinger is a leading international expert on... post was contributed by guest blogger Londa Schiebinger, PhD, Hinds Professor of History of Science, ... stem cells. In this case, they found that sex-matching of donor and recipient yielded the best results...the recipient of numerous prizes and awards. Schiebinger addressed the United Nations on the topic of ...