... advise pharmaceutical companies, biotech start-ups, health insurance companies, and governments on the...the part of the company involved: Research & Development (R&D) teams at pharma companies carry out preclinical... acquisitions of or collaborations with other companies. Medical Affairs groups work on building awareness... patients, shaping discussions with insurance companies about the pricing and coverage of drugs, and developing... teams, reflecting the need for all parts of a company to collaborate whenever it faces an important challenge...investments over the next 3, 5, or 10 years. Complicating these challenges is the fact that they occur...their jobs most, get pleasure from taking up such complex challenges, quickly breaking them up into smaller...

...derived from the more complex type III-A and III-B systems. Though it was composed of many protein subunits...might be phylogenetically classified as the more complex Class I systems that contain multiple subunits,...evolution, were also working on the project. Using computational observations, Koonin and Makarova found that...the domains into a single protein, shedding the complexity of its ancestors. They classified the system ...system contains two pieces: a single protein, computationally described as a fusion of four Cas7 proteins...significantly reduced amount of off-target toxicity compared to other RNA editing methods — streamlining experimental...mammalian cell lines. However, in those same comparisons, the AbuGoot lab reported no detectable effect...

... inquiry started when Neil Thompson, a research scientist at MIT’s Computer Science and Artificial Intelligence...technological change happens really fast,” Thompson says. To do this, Thompson reached out to MIT labs in the Synthetic...properties (GC content or rare codon percentage) and comparative properties (percent sequence identity and percent...query coverage) that could be determined through comparisons with a reference sequence database. Through a...content between many organisms and DNA synthesis companies often work within a narrow range for GC content...synthetic vs. natural gene are essential and complementary tools in a time where biological engineering...Kunjapur, Aditya M., Philipp Pfingstag, and Neil C. Thompson. "Gene synthesis allows biologists to source genes...

...Table 1: Taking it a Step Further - Big Company v. Small Company Big (e.g. Pharma) Small (e.g. Biotech...advancement in one company, good performers in for the long haul Limited movement in one company, likely will...academic silo to tenured professor.  Non-academia encompasses millions of choices including pharma, tech transfer...options in nonprofit science. No one list can ever encompass them all. We can’t designate non-academic jobs...and teach students Collaborations may be less “complicated” - for example, easy to initiate a collaboration...More sources of training and resources within the company Must find own sources of knowledge often externally...every 3-6 years—a short and exciting ride Complicated matrix structures are common Small integrated...

...when. Compiling the identity and location of a given nucleotide, a researcher (or a simple computer program...in Neuroscience at Harvard University. Just as computers, cell phones, and cars become more technologically... fluorescently labeled probe. If the DNA is complementary to the probe (for example, has the sequence ...be quantified by the strength of the signal in comparison to a control sample. Southern blotting applications...in standard PCR. This allows the researcher to compare different DNA sequences to see which is more abundant...mapping to investigate DNA sequence content by comparing the fragments created by different restriction... sequencing of many samples at once for later comparison. These next-generation sequencing techniques ...

...career at a biotech company, I always assumed I’d work one or two years at the company and then go to grad... the head of software product management at my company was open to meeting with me and explaining more...software product manager helps ensure that the company or software group is building the software tools...Well, I’ll give you an example. At my previous company, Ginkgo Bioworks, our software development team...backgrounds, most did not. It was my job to translate complex lab operations workflows into a format the developers... to solve is clear, but the solution is more complicated. You’ll need to be creative to be able to work...framework If you can’t find internships at science companies, try to find internships with translatable skills...

...protein:DNA complexes with antibodies specific to your protein(s) of interest. Once the complex is captured...protein:DNA complexes and finally (5) isolated the DNA from your antibody:protein:DNA complexes. Your DNA... the first step is to ensure your protein:DNA complexes will remain bound to each other until you are ...reversible bonds. This will keep the protein:DNA complexes together until you want to break them. Crosslinking...digestion can preserve more of the protein:DNA complexes. Once you’ve finished fragmenting the DNA, spin...that?  So while this method is certainly long and complex, you may be thinking, well, these are mostly fairly...techniques. What’s all the fuss? Well, ChIP’s complexities come not from the techniques used, but from ...

...requirement if you work for a private company you must show that your company employs at least 3 full-time people...Within these sectors, you could start your own company, work in a bench research career or a non-bench... to find the perfect job immediately after you complete your Ph.D. or postdoc. Foreign national researchers... need to be aware of the April 1 deadline for companies to submit an application on their behalf.  If ...technology transfer, communications, medical writing, compliance, core facility management, collaboration management...be directed to you specifically. You must also complete the peer review for it to be counted as proof ...your CV and bio sketch whenever you have new accomplishments to add. Obtain copies and translations of documents...

...engineered ESCs), and chimeric adults mated to produce completely transgenic offspring. Although powerful, this... genome, the CRISPR/Cas system makes use of 3 components, a guide RNA (gRNA) of about 125 nt that specifies...gRNA, Cas9, and donor oligonucleotide or plasmid components are brought together and microinjected into either...eggs. Or, to avoid handling embryos ex vivo, the components can be electroporated into the oviduct of pregnant... major advantages of CRISPR mouse editing when compared to more conventional methods. First, one can work... work with almost any strains of mice, as compared with conventional gene targeting, which is limited ...including 129 and C57BL/6, for which we have germline competent ESC lines. Second, the process is much quicker...

...MiSeq that takes about two days to complete. After the run completes, we begin our assembly process. Again...sufficient quantity for sequencing. This process is completed in a plate format yielding anywhere from two to...Illumina sequencing on our newly donated MiSeq.  For Complete Plasmid Sequencing we perform a 2x251 run on our...annotated Genbank files that are useful for more complicated plasmids, like those that contain synthesized...mutations, truncations, and insertions that could compromise function. When we do find mutations, we check..., NGS for some plasmids will not result in one complete, circular assembly. Most of the time, these plasmids...obtain an NGS result that is useful, but not 100% complete, we will still make this data available as a partial...

...ask your interviewers You’re interviewing the company as much as they are interviewing you, so come prepared...performance evaluations, professional development, and company culture.  It’s also a good idea to prepare 1-2 .... This is a chance to highlight your prior accomplishments and demonstrate your science communication ... Connect your prior work or a new idea to the company’s area of expertise. This has potential to make ...consider bringing your own laptop as a backup, if the company will allow it. Before you start your presentation...interview. Expect to be tired once the interview is complete Industry interviews are long! Mine lasted 4.5 ...read emails from HR carefully because sometimes companies have relaxed dress codes for interviews. You can...

...can we compare these methods in specific applications, we may also be able to use these comparisons to enhance...to 9 months in mice [2]). Low Immune Response: Compared to other viral vectors (e.g. Lentivirus, Adenovirus...is possible to use AAV constructs with self-complementing structure (scAAV) (7). As single stranded DNA...mapping of circuits In our lab, we use AAV-DREADD complexes to study neural function. Often the reporter proteins... proteins associated with these complexes are so robustly expressed that anatomical assessments can be...Ready-to-Use Retrograde AAV Preps at Addgene 2) Comparison of genetically engineered receptors (i.e. chemogenetics...neural control. Surprisingly, little research has compared and contrasted optogenetic and chemogenetic receptors...

...reaction or can be purchased 5’-modified from a company. However, unlike an RNA guide, it cannot be produced...probably while it is being synthesized) can form a complex with an ssDNA guide. Therefore, researchers must...inject cells with in vitro preassembled Cas9/gRNA complexes or with guide RNA- and Cas9-expression plasmids...survey and asked researchers if they had tested and compared NgAgo head-to-head with CRISPR-Cas9. The survey...respondents (transfection method, amount of each component per transfection etc.). Take Pooran's NgAgo Survey...same method used by Gao et al, 2016 (1). View the Complete and Up-to-Date Results of Pooran's NgAgo Survey..., leaving a small amount of active NgAgo/ssDNA complex in the cell. As proposed in the original article...

... function as a kind of “GPS for the genome” to compute coordinates for genetic elements like protein, ...action at a distance.” Through the marvel of DNA compaction, an enhancer can increase the expression of a...millions of base pairs away. (For details on DNA compaction and the structural proteins that make it possible...appendix DNA is a 3D fractal). Enhancer biology is a complex and dynamic field. We’re going to focus on a tried...People are wrong. In reality, cellular DNA is a complex three-dimensional globule of chromatin. Chromatin...tertiary structures (globules) to achieve exquisite compaction—on the order of 700 terabytes per gram—and form...chromosomes we all know and love. To understand compaction, suppose you have a piece string that’s 10 meters...

...library, as well as GC%, reverse compliment sequences, and more. Output_Compact: A text file that describes...there are also benefits gained by lowering the complexity of the screen. For example, the analysis becomes...a lasting impression on me. The slide showed a complex signaling pathway with many receptors, signaling...used these knockout mice to show exactly which components of the known pathway that were essential for ...be very useful for drug discovery, to dissect a complex signaling pathway, and to identify what part of...response to a particular ligand (such as when a component of a microbe or a cytokine is added to a cell)... rapidly performed by targeting all potential components of a signaling pathway. For example, one could...

...experts that complement your (scientific) skill set and is therefore very useful for complex projects. It...collaborations are becoming increasingly important as more complex approaches to scientific questions become popular...especially when considering small to mid-sized companies and organizations. Addgene as a nonprofit organization... worked with people outside the department to accomplish a variety of goals. For example, I worked with...provided a crash course in this discipline. Getting companies to come to these types of events can be very tough...First I needed to find the right contacts at the companies and then convince the contacts to give me some...repetition. I learned to look at the event from the company’s perspective, focus on the message and to keep ...

...challenging for sequences with complex secondary structure. Today, multiple companies offer long single-stranded...ribonucleoprotein (RNP) complexes of Cas9 protein and the separated guide RNA components (crRNA and tracrRNA...maximum Easi-CRISPR efficiency. Easi-CRISPR is compatible with standard microinjection and electroporation...successfully used guides targeting the same or complementary strands, as well as guides facing towards or... simple method for delivery of genome editing components to the germline. In contrast to standard microinjection...placed to mediate electroporation. Ohtsuka et al. compared traditional microinjection and i-GONAD knock-in...efficiency using a small ssDNA, and they found comparable efficiencies (52 vs 49%). In contrast, Easi-CRISPR...

...with fluorescent proteins (FPs) localized to compartments other than the cytoplasm. That is, unfortunately...positioning of the FP sequence relative to cell compartment targeting sequences, which often have absolute...that FPs are often poorly suited for cellular compartments other than the cytoplasm. Generally, FPs evolved...subcellular environments, including the organelles that compose the secretory pathway, endocytic vesicles, mitochondria.... Many of the resident proteins within these compartments undergo significant post-translational modifications...inter-chain disulfide bonds between FPs blocks Golgi complex-localized fusions from properly exiting the ER ...and spectral characteristics of the moxFPs are comparable to the non-optimized parental proteins. Our recent...

...GPCRs (Takeda S et al., 2002) and many downstream complexes involved, it can be difficult for scientists to...in sight (Olsen RHJ et al., 2020). A transducer complex consists of a trimer of three subunits: alpha, ...collectively known as a heterotrimeric G protein. This complex converts a signal received by a GPCR on the cell...is being able to identify specific transducer complexes that are activated in response to a ligand binding...inactive, these subunits are found in a trimeric complex bound to the intracellular surface of the GPCR....Following activation in response to the ligand, the complex will disassociate, resulting in a decreased GFP2...They further used TRUPATH to analyze a panel of compounds across the transducerome on a single GPCR, showing...

...effects, ABEs also compare favorably to other methods. In a head to head comparison with Cas9, Cas9 modified... efficiency of homology-directed repair (HDR). Compared to non-homologous end joining, HDR occurs at a...represent the fourth generation of base editors. Compared to BE3, BE4 offers a 2.3 fold decrease in C->G...frequency of 1.2%. Although the lab did not conduct comprehensive genome-wide studies of ABE specificity, their... 40 new ABE8 variants (Gaudelli et al., 2020). Compared to ABE7.10, ABE8s resulted in 1.5-fold more editing...higher editing efficiency at non-NGG PAM variants compared to ABE7.10. ABE8s have an improved base editing...done by fusing the adenine and cytosine editing components together. Keith Joung’s lab created a dual-deaminase...