... tactics of prokaryotes to deliver the technological biological blockbuster known as the CRISPR (clustered...genes, which code for polymerases, nucleases, and helicases, were also disrupted to elucidate their role in...destroy the phage upon re-exposure (2, 3). More specifically, specialized Cas proteins snip the foreign DNA...distinct types and subtypes using polythetic classification based on evolutionary similarity (5). Type ...2013 (7, 8). Nine months later, more than 1500 publications describe work to improve the tool’s specificity...various species and describe an assortment of applications. Despite the ubiquity of CRISPR/Cas toolkit,...programmable platform well-suited for various innovative biological and translative technologies. Addgene has empowered...

...lab while the publication page contains all the plasmids deposited from the same publication. Next up, we... Addgene’s plasmids is an indication of popularity. A yellow flame indicates that a plasmid has been ordered...through the depositing lab page, then onto the publication page, and finally on down to the individual plasmid...Great! Now, if I want to use this plasmid in a publication, how do I know who to acknowledge? Depositing...plasmids that are available through Addgene. Publication: If the plasmid has been published, the paper...the bottom of the page that help you cite this publication in your own work: How to cite this plasmid: The...depositing lab whenever their plasmids are used in a publication. Addgene makes this easy by providing a template...

...methods. Since molecular, ecological, and reproductive confinement typically require very little effort...Development Fellow at the Wyss Institute and Harvard Medical School. Scientists making transgenic organisms ...the entire wild population with unpredictable ecological effects. Because the consequences of such a mistake... to implement and have been tested in yeast5. Ecological confinement involves performing experiments in...experiments attempting to build gene drives in tropical mosquito vectors of diseases such as malaria and...they should not be relied on exclusively because historical studies of pathogen research have conclusively...strategies is strongly advised. The effects are multiplicative, and there are very few gene drive experiments...

...Castano Corsino, a junior faculty member at Harvard Medical School/Brigham and Women’s Hospital (BWH). Based...like IMP, bring closer than ever (although not physically) the ideas and experience from talented professionals... IMP USA-Europe not only provide the specific technical and logistic support to increase the chances of...professional opportunities that are outside of our typical boundaries, and will arrive to Spain at one point...international mentors. Universities from five different geographical locations in Spain were enrolled: the University...represented a diverse group of individuals with technical and career path needs, and came from the universities...that their knowledge and skills had improved significantly when compared to their pre-program self-assesment...

... editing in Escherichia coli. Acta Biochimica et Biophysica Sinica. https://doi.org/10.1093/abbs/gmab036...for successful modifications, efficiency can be low, especially for larger modifications. What’s the solution...tools available for bacteria and some of the applications for which they’ve been used. The beginnings ... of Enterobacteriaceae, without the need for modification. This finding suggests that the system may be...eukaryotes (Qi et al., 2013). As in other systems, catalytically dead (dCas9) targeted to a promoter or gene ...gene body can repress transcription by physically blocking the elongation complex from binding the DNA or...responsive promoter to drive expression of a catalytically dead Cas9 enzyme allows reversible repression...

...where you assume your technical skills define you and that the other not-so-technical skills you’ve built...my job. The same can be said about teamwork, communication, writing, management, and creativity; I have... individual with more going for you than your technical expertise in a specific field. While searching...built over time are only applicable to those positions you have already held. Imagine this situation: You...have the desired degree, you have mastered the applications they use, you’ve gained experience working with...under your belt from your last job. You knock the technical portion of the interview out of the park. It’s...hiring team will also want to examine not just technical skills, but also transferable skills, before offering...

...Cell-Type-Specific Gene Editing." Journal of the American Chemical Society 140.21 (2018): 6596-6603. PubMed PMID...valuable to scientists, there are some potential complications that must be considered when using this method...alternative approach, which avoids many of these complications, is to directly deliver a ribonucleoprotein ...used for most of the current genome engineering applications of CRISPR including: generating single or multi-gene...pathways. This makes Cas9 RNPs useful for CRISPR applications where limited expression of Cas9 is required... of eventually using this method for in vivo applications. This technique utilizes Cas9 proteins harboring...Liang et al. 2017).  PEG-mediated methods, while technically less difficult to perform, often show low efficiency...

...mismatches on modification by AsCpf1 (red) or LbCpf1 (blue). Mismatch positions are indicated below each ... this publication at Addgene. Makarova, Kira S., et al. "An updated evolutionary classification of CRISPR-Cas...it lacks Cas9’s other HNH endonuclease domain, indicating that Cpf1 functions differently. Since Cpf1 loci... Direct synthesis of these crRNAs should be significantly cheaper than that of the ~100 nt crRNA/tracrRNA... off-target editing rates, determined using sophisticated genome-wide analysis. At many of its computationally...compared the ratio of total off-target to on-target modification for AsCpf1 and LbCpf1, and found that both orthologs...-target editing in a cell culture model. The application of Cpf1 to genome editing is exciting both in...

...require grant seekers to use online applications with significant character limits for the written portion...development, certification, degrees, foundations, coaching and even Twitter Chats dedicated to the Grant...the candidate application process, the candidate’s ability to write a grant application through a written... much more than writing. In fact, many grant applications and grant makers have moved away from long written...portion of the grant application. They have increased their demand for stronger budgets with clear budget...duties can range from coordinating the grant application and submitting it, to evaluating the capacity...especially in the sciences and by extension to scientifically oriented nonprofits. Like Deb, I worked for...

...dynamic, physiological processes. Re-engineering GFP to increase its range of color and application Almost...through mutagenesis in order to improve its physical and biochemical properties. In 1995, Roger Y. Tsien described...transgenic mice. Purification: GFP can be used as a general epitope tag for protein purification and a number...scientists to begin to study GFP and deduce its biochemical properties. Now GFP and its fluorescent derivatives...GFP is used in research across a vast array of biological disciplines and scientists employ GFP for a wide...their experimental needs, and some of the common applications in the lab. Why Green Fluorescent Protein?...others) Neutral mutations    A multitude of applications Due to its size and ease of use, GFP and other...

...after subtracting out non-specific binding). Historically, glutathione-S-transferase (GST) fusion proteins...advantage of a fusion protein is the efficient purification of an already immobilized target protein. The...immobilization method. One possibility is covalent modification of the purified target protein with the vitamin...derivatives streptavidin and neutravidin) is for all practical purposes covalent. Therefore, protein immobilization... four biotin-binding sites per molecule so, theoretically, saturated avidin could form tetramers of biotinylated...well-controlled. The lack of site selectivity typically diminishes the apparent affinity for the ligand...However, large amounts of the target protein are typically required due to the relatively low efficiency ...

...genomic modifications, is very robust in C. elegans, and the methods for HDR-mediated modification continue...permission from the Genetics Society of America. Applications of this system Dickinson et al. have developed...Used with permission from the Genetics Society of America. To use this SEC-based system, first design...use and highly efficient for C. elegans genome modification. Figure 2: Schematic from Dickinson et...system could be used to make many other genome modifications, including targeted mutations. SEC-based systems... in cell culture. In addition to CRISPR/Cas9 applications, SEC selection could replace other, more restrictive... of print] PubMed. Find plasmids from this publication at Addgene. Other strategies for CRISPR/Cas9...

...mammalian cells often relied on chemical (CaCl2, cationic liposome) or electrical (electric current forming ...research since their first use in the early 90’s to genetically modify primary cells. Amongst the different vectors...particles and the virus particles produced could not replicate. However, Mother Nature found a way to re-create...packaging plasmid forming a vector that could create replication competent virus. Even though this system was ...its safety was a disadvantage particularly for clinical development. In addition, MoMLV is inefficient...still a potential risk for the generation of replication competent lentiviruses (RCL) especially if you...factors in HIV positive cells could make the virus replication competent. The 3rd generation of lentiviral...

...post was updated on Jul 27, 2020. CRISPR, and specifically Cas9 from S. pyogenes (SpCas9), is truly an ...use, functional in most species, and has many applications. That said, SpCas9 is not the only game in town... makes this molecule unique, and the various applications of Cas13a. The origins of Cas13a: An RNA cleaving...specified by the crRNA sequence, it adopts an enzymatically “active” state rather than reverting to an inactive...discussed below. Subscribe to CRISPR Blog Posts Applications of Cas13a   Figure 1: Using Cas13a as...) is amplified using Recombinase Polymerase Amplification (RPA) or Reverse-transcription RPA (RT-RPA),...fluorophore. The fluorescent signal can be used as an indicator to determine whether the target sequence is present...

...tropism, replication kinetics, and cell damage profiling of SARS-CoV-2 and SARS-CoV with implications for ... from their original sources. Figure 1: Biological repositories that distribute mouse strains, plasmids... plasmids, cell lines, and much more have been critical for pandemic research. Image from Reproducibility...this virus. One such example is cell lines. The American Type Culture Collection (ATCC) is a private, nonprofit...Cell Bank, part of the National Institutes of Biomedical Innovation, Health, and Nutrition in Japan. These... Another example of reagents that have broad applications but have proven useful to the study of SARS-...vectors VSV-G and psPAX2, which were used in a publication by Xiuyuan Ou, et al. that characterized the ...

...overall. Communication is the issue Much of the problem can be attributed to communication, and in particular...particular communication about experimental methods and approaches. It’s very easy to think that the methods...that they were doing something differently. Communication comes in many forms—from transparency about ...them troubleshoot and understand. All of this communication and sharing fits together to make for a transparent...for pushing for more transparency in science communication falls on the shoulders of journals and publishers...can feel like yet another hurdle in front of publication when all they really want to do is get the paper...we take our responsibility to push for clear communication of science very seriously. That’s why at Cell...

...skills The ability to think critically and carry out in-depth research is applicable to a wide range of industries...many opportunities available to those who are scientifically minded! Listen to Biotech Founder Michael Koeris...Scientific Advisor Lab Manager Patent Lawyer Technical Writer Meteorologist Journal Editor Textile ...organised, reliable, and can work independently. Practical outdoor skills are needed in a variety of roles...Breeder Scientific Illustrator Plant Breeder Zoological Worker Conservationist Documentary Director ...range of roles, including: Science Reporter Technical Writer Statistician Bioinformatician Science...Marketer Software Developer Learn Science Communication Skill from a Student Group Like Science in the...

...target antigen as canonical antibodies. They also have the advantage of being significantly smaller, meaning...binding to cell surface receptors. These applications are typically used for investigating signaling pathways...tool that can be used for a variety of research applications. Within the immune system, antibodies are produced...antibodies to bind proteins is useful for research applications as well because they allow scientists to target...Epitopes are between 5-8 amino acids long, with a typical length of 5 or 6 amino acids (Cruse et al., 2004...recognize epitopes that full antibodies would not physically be able to reach. While a full antibody is ~150...150 kDa, a nanobody is 12-15 kDa.   Research applications for antibodies You can use antibodies for experiments...

...cells and statistically analyze cell movements. By using cells labelled by fluorescent chemicals or by other... steady state or under physio-pathological challenge. However technical specificities have to be taken... required for a growing organism to make the complicated transition from a clump of cells to a fully differentiated...mobility than the static microscopy images of chemically coloured lymphoid organs did in the past. How...fluorescent protein or dye. It separates the cells mechanically in a vibrating nozzle, imparting a positive ... millimeter in depth. Two low-energy photons (typically from the same laser) cooperate to cause a higher-energy...reporter constructs in the immune lineage. By specifically labeling T cells, B cells, and antigen presenting...

...colleges, private universities, and medical schools. I will emphatically deny that I have a magic formula..." manual on applying for faculty positions in biomedical science. My motivation was to share my experiences... do need multiple first/corresponding author publications in quality journals. It helps to come from the...a job. The currency of scientific success is publications and grants. 2. Write succinct and easy to understand...told all of the nuances of the research in as technical of language as possible. Seriously. My suggested...humbling. My conviction that I had a flair for communication was rapidly dispelled. She would ask me, "Is... trying to say X." She would then look at me quizzically and ask why I didn't just say that? Bottom line...