...two different pairs of chromosomes using gRNAs specific to sequences on chromosomes 9 and 13. They next ...example, the visualization of chromosomal translocations or chromosomal shattering (chromothripsis). As...catalytically dead Cas9 (dCas9) has many applications, the most well-known being transcriptional activation/repression...could then be used to examine both inter- and intrachromosomal dynamics, and how they change in response ...turned their attention to mapping pairs of intrachromosomal loci. The technique successfully resolved ... this work represents the first mapping of intrachromosomal loci, a major benchmark in characterizing ...next! References Multicolor CRISPR labeling of chromosomal loci in human cells. Ma H, Naseri A, Reyes-Gutierrez...

... biosensors for Human Milk Oligosaccharides (HMOs). HMOs are thought to play an important role in nuturing...to engineer bacteria for the efficient product of HMOs that could be used to supplement newborn diets and... cost-effective way to measure the production of HMOs. Current HPLC-MS and fluorophore-based technologies...biosensors that could quickly and reliably detect HMOs. Fatima’s first design is a whole cell biosensor...specifically proposes using this sensor to detect HMOs found on cellular surfaces. After incubating the...the sensor with bacteria engineered to produce HMOs, sensor binding results in an increase in fluorescence...fluorescence of cells that actively produce the HMOs. Fatima has demonstrated the successful production of both...

...when I recently attended the 2018 Chromosome Architecture and Chromosome Organization Keystone Symposium...nucleosomes, loops, chromosomal domains, or how stretches of DNA on different chromosomes interact with each...posters I saw at the 2018 Keystone Chromosome Architecture and Chromosome Organization. These aren’t the ...linear” is a misnomer. The DNA packed into our chromosomes is very much 3-dimensional, as you can see by...Shapes a Single Nucleus from a Set of Mitotic Chromosomes. Samwer M, Schneider MWG, Hoefler R, Schmalhorst...

...resistance cassettes is most commonly employed, with the KanMX being the most prevalent due to its versility...highly versatile DNA transformation system.Unlike most other microorganisms, yeast have both a stable haploid...and must be integrated directly into the host chromosome via homologous recombination. Yeast Replicating...Replicating Sequence (ARS) derived from the yeast chromosome. As the name suggests, these vectors can replicate...replicate independently of the yeast chromosome; however, they tend to be unstable and may be lost during...replicate as though they are small independent chromosomes and are thus typically found as a single copy...integration. Yeast Episomal plasmids (YEp): These are most similar to bacterial plasmids and are considered...

... but almost all of these guidelines can apply to any workplace or job. It amazes me when the most analytic...”  and hone in on the factors that will make the most difference for your success. Gather data on the ...if they are not local. Alumni meetings can be the most informative way of gathering data. Former lab members...totem pole). Can you see yourself enjoying this atmosphere? Lab size matters Yes, yes it’s all about ...the science but really it’s not. It’s about the atmosphere that will allow you to thrive. How big is the... the energy level in the lab? Does a frenetic atmosphere make you edgy or energized?   Are you an only...backbone”. The sad truth is, if it happened before, it almost certainly will also happen to you. As you speak...

...integrations on the genome. Most recently, we’ve developed a new system “Multicopy chromosomal integration using...bacteria. Our pCas/pTargetF system has been one of the most popular genomic editing systems in E. coli. However... both plasmids into the cell. The edit in the chromosome can now take place. After editing, pTargetF can...great for experiments that require only single chromosomal integration (ex: genome editing), our MUCICAT...transposase system, INTEGRATE, is best for multicopy chromosomal integration (ex: for protein expression) (Zhang...Zhang et al., 2020). Multicopy chromosomal integration for gene expression is an alternative versus plasmid-based...advantages: Fast and site-specific multicopy chromosomal integration of cargo without using screening ...

...There’s an astounding number of ways to create chromosomal mutations in bacteria, so many that it may be...quick and easy way to introduce a mutation in the chromosome is to disrupt expression of a gene with an antibiotic...resistance cassette. This leaves a “scar” in the chromosome, sometimes interfering with expression of surrounding...ones that don’t leave any undesired scars in the chromosome. These methods roughly fall into two categories...domesticated from the Lambda bacteriophage, is arguably the most widely and productively used  (Thomason et al., ...introduced from a conjugative plasmid replaces the chromosomal copy. This is a versatile technique, and is less... markers used to identify double crossovers? The most commonly used counter-selection system relies on...

...term familiar to almost every person regardless of profession, race, or gender. Almost no other disease...material is damaged beyond repair, that cell will most often commit itself to the apoptotic pathway and...eliminate cancerous cells from the body. One of the most prominent of these is the Ras pathway. The term ...exploiting the different mutant versions of Ras that are most prevalent in human cancers, and synthesizing compounds...mutated residues. For Ras researchers, one of the most exciting and relatively new resources is the RAS...Further, for each plasmid in the kit, the transcript most commonly found across all TCGA tumor samples is ...New resources and technologies become available almost daily, yet obstacles remain. One of the biggest...

...of recombinant protein expression and some of the most popular commercial strains designed to avoid them...usage (Table 1) A few mutations are common to all or most expression strains to accommodate high protein levels...  The DE3 lysogen/T7 promoter combination is the most popular induction system. The DE3 lysogen expresses.... Although the DE3/T7 RNAP system works well for most experiments, the lac promoter can “leak,” meaning...exists even without the addition of IPTG. This is mostly a problem for toxic protein products, which can...should generate sufficient expression levels for most purposes, but what do you do when you’ve tried a...solubility concerns. Insoluble and Secreted Proteins: The most common purification protocols are designed for soluble...

...healthcare consultant, I have worked on most of these kinds of projects. Almost all of my projects, have involved...health insurance companies, and governments on the most effective and efficient ways of delivering healthcare...that specialize in business strategy tend to be mostly engaged by pharma and biotech clients. In this ...Who would enjoy healthcare consulting? One of the most rewarding parts of the job is being confronted with...healthcare space. The consultants who enjoy their jobs most, get pleasure from taking up such complex challenges...situations can get stressful and to help manage that, most consulting firms coach and counsel employees on ...of the day, the consultants who enjoy their jobs most are those who understand that these challenges are...

...making it unsuitable for most in vivo applications.  Third, even in the most optimized tissue-culture ...sequence-specific knockins - for which the AAV approach is most well suited, since it routinely generates knockin...this is comparable or better than the efficiency of most current CRISPR-based approaches for the generation...several recent reviews (Howes and Schofield, 2015; Moser and Hirsch, 2016; Epstein and Schaffer, 2017) Hopefully...Jung-Sik, et al. "Intact cohesion, anaphase, and chromosome segregation in human cells harboring tumor-derived...engineering using adeno-associated virus (AAV)." Chromosomal Mutagenesis. Humana Press, New York, NY, 2015...2015. 75-103. PubMed PMID: 25408402. 9. J Moser, Rachel, and Matthew L Hirsch. "AAV vectorization of DSB-mediated...

...still not get the most from your team. It is pretty clear that if people are mostly happy at work, they... and grow in skills. Managing a team is hard and most people have some learning they need to do before...overwhelmingly agree that these are the factors that most contribute to job satisfaction: A feeling of being... not mean a manager must allow his team to work mostly at home. Having a high degree of freedom and diversity...pursue and problems to solve (I think this is true of most people, not just scientists) Are self-directed, ...be in touch with the needs of the team and to be most effective in their role. The best managers have ...

...this useful. Now that we’ve been doing this for almost 6 months, you may be looking for more ways to make...don’t all get the mentoring support we need, and most of us could use more mentors to listen, guide and... virtual!   2. Practice public speaking For almost any science career, there will be a time when you... about trying your hand at a video presentation? Most video platforms make it easy to record yourself....audiences in your future.     3. Start something In almost every career talk I present, I’m asked what Addgene... different criteria for different roles but for almost every position, we look for candidates that have... interviews, for more details on how to make the most of these exploratory conversations. If you set yourself...

...the emission of photons. The most familiar bioluminescent organism for most people is the firefly (Photinus...pyralis) and perhaps not surprisingly it is also the most commonly used bioluminescent reporter. This beetle...from the firefly beetle (Photinus pyralis) is the most frequently used bioluminescent reporter, other luciferases...substrate coelenterazine and is likely the second most commonly used luciferase. Other organisms from which...assay The typical luciferase-containing plasmid is most often used to investigate the effect of regulatory...Many luciferase assays require cell lysis as the most efficient means to disrupt the cell membrane and...

...which chromosome the transgene has integrated and make genetic crosses with balancer chromosomes to make...that the transgenes can be inherited on the same chromosome. Targeted Insertion allows for direct comparisons...longer because you need to determine in which chromosome your transgene has integrated. This can be accomplished...accomplished with a few matings to balancer chromosome lines but is not necessary with targeted insertions...insertions (here you would know beforehand on which chromosome your docking site is located). This amounts to...genomic locus of integration is. The white gene is most often used, but other selection markers include ...

...There are a lot of viruses out there – most of which we would all like to steer clear of! However, a ...them a little better. Here we will discuss the four most commonly used lab viruses: gamma-retrovirus, lentivirus...host's genome and is used to make more virus. The most studied (and used in the lab) type of gamma-retrovirus... have a cargo capacity of ~ 8 kb, ample room for most experimental needs, and can transduce (infect) a...integration, and its association with disease. The most well-studied lentivirus is HIV, which many lentiviral...replicate episomally, essentially remaining an extrachromosomal part of the host genome that can replicate... However, the cargo capacity of AAV is ~4.7 kb, almost half that of lenti and gamma-retro viruses. This...

...also has the flexibility to modify the E. coli chromosome, plasmid DNA or BAC DNA.  To use the lambda ....   This setup can be also be used to modify chromosomal genes. After the initial editing event, the defective... defective prophage can be removed from the chromosome of the host E. coli by a second lambda red recombination... system is best suited for editing bacterial chromosomal DNA because it is easy to remove the recombineering...responsible for excision of mini-λ from the host’s chromosome. After this, mini-λ can readily be purified from...PCR can be used to screen for positive clones in most cases, and restriction enzyme digest can be used... being targeted for modification: the E. coli chromosome, a plasmid, or a BAC.    Pro Tip: ssDNA substrates...

...your lab, don’t be surprised. They are present in most of the cell culture facilities, tissue culture labs...is difficult to be detected in the serum that is most commonly used for every cell culture. Reusing the...agar plates. This method is capable of detecting most of the mycoplasma species, apart from a few of them... are used. A broad range of primers encompasses almost all the infecting mycoplasma. The primers used ...Prevention of mycoplasma contamination The best and the most important ways to avoid mycoplasma are by providing.... Plasmocin is a widely used drug that can clear most of the mycoplasmas present in the culture media.... MRAs are non-toxic, effective in the removal of most of the mycoplasma contamination, easy to use, and...

... skill in its most difficult form, which makes it the perfect transferable skill. Almost every other form...Public speaking is not just one of the most common phobias–it’s a key part of being a scientist and an... ideas clearly and strategically is powerful in almost any career. It allows you to share and build support...and take a deep breath before the panic sets in. Most of the time, that’s enough–but if it’s not, I can...memorable and awful. So try to keep some perspective. Most people won't notice, or won't remember and those...is that you will have to learn this skill in its most difficult form. You’re presenting complex data, ...

... types of viruses generated in the lab. The four most common types of viruses grown in the lab are gamma...infection process (though it can integrate at chromosome 19q13.4 qtr. (AAVS1)). For these reasons, AAV...to work with BSL-2 materials, such as lentivirus. Most of this work will need to be done in a biosafety...a significant risk for aerosol contamination. As most centrifugation has to be done outside of a biosafety... and scrapes from sharp equipment are one of the most common forms of lab accidents, so extra precaution...significantly reduce the risk of an accident (the most common reason for lab accidents is not following...corners. For many researchers, viruses might be the most dangerous things they work with, and that can understandably...