... you have a genome with unknown sequence. In the era before sequencing was commonplace, researchers used... ultimately leading to what are generally considered first generation DNA sequencing machines (9). Today... amount of high-quality DNA, real-time PCR has several advantages including easier automation, higher ...both time and resources (3). Still, researchers generating transgenic animals may choose Southern blotting.... Today, researchers use both Sanger and next-generation sequencing techniques to answer similar experimental... subset to sequence. Sequencing gels and next-generation sequencing techniques Introduced in 1977, Sanger...samples at once for later comparison. These next-generation sequencing techniques do not typically rely on...

.... Why an International Mentorship Program? In an era where millions of people hold a PhD, achieving our...people and society. Future generations of IMP Motivating future generations is key to the success of this...world, professional internationalization faces several obstacles; for instance there are few resources...Programs at BWH and providing career mentoring to several students from Spain and all over the world, she...the country. Brain drain is a serious problem in several countries, however programs like IMP, bring closer...from international financial struggle, our new generations feel thrown, to sink or swim on their own, into...Development, and Innovation, D. Carmen Vela, who reiterated the Spanish government’s support for the IMP ...

...threshold of a revolution in modern neuroscience, an era in which technologies like FLEx and optogenetics ...arousal in mice previously subjected to steady-state general anesthesia (Taylor et al., 2016). These very same...modulation of FLEx vectors combined with the use of several Cre transgenic mice targeting different brain areas...PMCID: PMC6317997. Deisseroth, Karl, et al. "Next-generation optical technologies for illuminating genetically...exceeding the carrying capacity of AAV vectors." Gene Therapy for Neurological Disorders. Humana Press, New York...26135556. Hudry, Eloise, and Luk H. Vandenberghe. "Therapeutic AAV gene transfer to the nervous system: a clinical...ventral tegmental area induces reanimation from general anesthesia." Proceedings of the National Academy...

...technologies need to be discarded but clearly we are in an era of neuroscience research where optogenetics will ...methodological and experimental limitations and considerations? I recently spent two months in the lab of ... at NIDA and some of the practicalities and considerations you may not find in the materials and methods...prove that your optogenetic manipulations are generating action potentials (or lack there of, depending...hyperpolarization, and inhibition of action potential generation. Archaerhodopsin (Arch) is activated at a similar... no longer available through Kopf). Additional general protocols on stereotaxic surgeries for viral injections...are obviously limited by the mouse line itself. Generation, breeding, and maintenance of a mouse line is...

...Existing therapies and effects on immune system Conventional cancer therapies include radiation therapy (owing... Types of immunotherapeutics In lieu of the cancer-immune system interactions, therapeutics have been ...Curie), chemotherapy, and surgical removal of the tumor. However, these conventional therapies produce...cell-surface tolerance-producing proteins are one of the most commonly used immunotherapeutic agents. Currently...contestants for cell-based immunotherapy. Using activated DCs ex vivo to generate cytotoxic effector T cells...beneficiaries. Collaborative therapy For the most efficient outcomes, collaborative therapies must be explored. ... multiple immunotherapy targets can be utilized at the same time. A combination of tolerance marker blockade...

...combination of the promoter (RNA polymerase binding site) and operators (response elements). Promoters ...      The RNA polymerase(s) RNA is transcribed from DNA using an RNA polymerase (RNAP). In bacteria...     Description Expression Additional considerations CMV General expression mRNA Strong mammalian expression...PGK1 (human or mouse) General expression mRNA Mammalian promoter from phosphoglycerate kinase gene.  Constitutive...promoter with low basal activity and several tetracycline operators. Transcription can be turned on or ...promoter. GDS General expression mRNA  Strong yeast expression promoter from glyceraldehyde 3-phosphage ...Description Expression Additional considerations T7 in vitro transcription/ general expression Promoter from ...

...under the control of the cI857 temperature-sensitive repressor. The temperature-controlled expression of the...allows genome editing and mutant library generation in several biotechnologically and clinically relevant...proteins. Overall BioID2 improves the efficiency of screening for protein–protein interactions. The Roux...on protein-protein interactions and therefore identifying and studying interaction networks is important...Protein A-Renilla luciferase and the “prey” proteins are tagged with Firefly luciferase. The bait and prey...A tag. The luciferase activities can then be measured and normalized to assess interactions. Higher relative...firefly luciferase activity in the Co-IP compared of a control Co-IP indicates stronger interaction. The ...

...Current CRISPR Therapies Cancer and CAR-T therapy The FDA has already approved CAR-T immunotherapy for cancer...delivery and editing strategies for CRISPR therapies. Systemic therapies involve CRISPR-Cas delivery directly...next few months. Several other companies have followed suite, with Beam Therapeutics initiating a base... initiated a CRISPR therapy for hATTR with the first-ever systemic CRISPR therapy. They engineered a CRISPR-Cas...this therapy is affecting hATTR patient outcomes. Bacterial infection and HIV Most CRISPR therapies have...part of a cocktail therapy for urinary tract infections. The Cas3 enzymes target several strains of E. coli...sponsored by Excision Biotherapeutics. Additional ongoing trials The CRISPR therapies don’t stop with the...

...spatial and hierarchical chromatin interaction networks. Think of these chromatin interactions networks ...pictures” generated with current 3C methods provide useful information about chromatin interactions, they ...could detect multiple chromatin interactions at once. Chromatin interactions at five sites in the well-characterized...chromatin interactions identified with the multiplexed CAPTURE 2.0 largely replicated interactions previously...Determining the spatial and hierarchical organization of chromatin interaction networks  A second application...enhancers are interacting with which promoters, what part of a gene enhancers like to interact with, and ...that forms chromosomes. These long range DNA interactions are often detected with chromosome conformation...

... that the ligation regenerates the original restriction sites for further iterative cloning steps and ...and voltage sensitivity. This hierarchical screen led to the generation of two enhanced voltage indicators...Instead, coelenterazine is supplied to the culture media as a chemical substrate for Renilla luciferase. This...alternative to the widely used luciferase reporter gene plasmid pGL3 pGL3 luciferase reporter gene plasmids... issues with fluorescent pH sensors, a set of luciferase-fluorophore pH fusion reporters, pRSETb-pHlash...insensitivity to other ions In BRET, A mutant Renilla luciferase, catalyzes the oxidation of its substrate, luciferin...control of signalling pathways. These LIDs are generalizable, versatile and powerful tools which allow you...