...models for developmental processes and promising therapeutic tools. The two unique characteristics that define...differentiate into specialized cell types [2]. We general speak of 4 levels of stem cell potency: Totipotency...They are isolated mostly from abnormal and/or supernumerary preimplantation embryos that are produced during... in many organs in the adult human body. These generate specialized cells for their location, mostly to... bone marrow only differentiate into cells of peripheral blood. Typically, there is a very small number...cells can be used for different research and therapeutic purposes. Both types have their advantages and...isolated from the human body Usage Universal. Can generate all adult cell types. Limited. Only specific cell...

...possibilities for the clinical and therapeutic applications of iPSCs. The generation of human iPSCs (hiPSCs) now...programming spanning from fibroblasts and keratinocytes to peripheral blood mononuclear cells (PBMCs). PBMCs...LMO2-associated clonal T cell proliferation in two patients after gene therapy for SCID-X1. Science, 2003... Fukuda, Generation of induced pluripotent stem cells from a small amount of human peripheral blood using... to generate integration-free human-induced pluripotent stem cells from cord blood and peripheral blood...(Kushi) Mukherjee, a Postdoc at Massachussetts General Hospital. A little over a decade back when Yamanaka...Yamanaka”) factors, they changed the landscape of regenerative medicine. Their work opened up vast possibilities...

... it inspired. CAUTION: Spoilers alert! Pre-DNA Era  H.G. Wells, the father of scientific fiction, was... is now famed as the Griffith’s experiment.  DNA Era The period between 1940-1960 was pivotal for genetics...authors during this period.  Genetic Engineering Era: Pre-CRISPR  Having visualized and understood the...preference than the ZFNs.   Genetic Engineering Era: Post CRISPR  CRISPR The strategy of using a protein...molecular genetics was booming, SciFi saw the rise of literary giants such as Isaac Asimov, Arthur C. Clarke,...nucleases (ZFNs) are artificial restriction enzymes generated by fusing a zinc finger DNA-binding domain to ...finger recognizes a nucleotide triplet and when several ZFNs are strung together, it can read many base...

...Potential applications for gene and cell-based therapies in the new era of personalized medicine (2017). ...Potential applications for gene and cell-based therapies in the new era of personalized medicine." Genes & diseases...nucleocapsid. B) Three generations of adenoviral vectors: the first generation vectors are deficient in...in trans. The Gutless AdV have generated high interest for gene therapy due their increased cloning capacity...attractive vectors for research applications and gene therapy: they can be produced at high titers, can accommodate...the Adv genome’s utility in research and its therapeutic potential. Note: constructs discussed are based...based on Adenovirus type 5. Three generations of adenoviral vectors The adenovirus virion is a non-enveloped...

... trusting science in a “fake news and post-truth era,” the politicization of science (from climate change...intellectual prowess and scientific skills. This is an era of social media and self-promotion, and personality...contributed to the steadily increasing distrust the general public has in science and scientists (as discussed...have addressed CRISPR frequently over the last several years, but lay readers may be less familiar with... interesting case studies, and Zimmer provides several. In Part Three of the book, called simply “Old ...

..., a chalk talk is quite simple. It's you, a dry erase board, some markers (chalk is rare these days), ...of your chalk talk Bring your own fresh set of erasable markers. You’ll have the colors you want and you...in 1-5 years, and contribute to resolving the overarching knowledge gap. Aims should have short titles,.../goal, an approach, anticipated outcomes, and preferably a description of the payoff if the aim is successful...bioengineered scaffolds with patient stem cells to generate compatible functional livers.” Why are you the...getting back to your story. Committee questions can derail your talk. You need to balance respectfully answering... for presenting a science talk Check out our transferable skills guide Read all of our career blog posts...

...an indel or inversion mutation [3]. In the CRISPR era, the genomic landscape may be surveyed in unprecedented...present a video reprint of the CRISPR article "Generation of Genomic Deletions in Mammalian Cell Lines ...between deletion size and frequency [3]. There are several applications in which the production of deletions...within the deletion boundaries may be produced to generate clonal allelic series. Single DSBs resolved by...

...colleagues can be more difficult during the pandemic era, with fewer opportunities for spontaneous conversations...creative interaction During Zoom meetings, different tools can be used to promote creative interaction. A great...here, and only general chatter will be allowed with a different host each time that moderates conversations...An example could be making lab meetings more interactive and creating space for discussion at the end ...end of each talk - time for free interaction and brainstorming. This could be achieved by asking each presenter...also make the screen-sharing experience more interactive. The button is on the right of the Share Screen...writing publications/grants too. As described by several works (Wuchty et al., 2007, Vicens et al., 2007...

...methods for C. elegans were released, signaling a new era in C. elegans research. Homology directed repair ...CRISPR in C. elegans even easier - now, one can generate a fluorescent protein fusion, transcriptional ...efficient selection-based CRISPR/Cas9 method to generate knock-ins on any genetic background. Their new...flanking loxP sites and a heat-shock inducible Cre to generate a self-excising cassette (SEC). To make it possible...

...Potential Applications for Gene and Cell-Based Therapies in the New Era of Personalized Medicine. Genes Dis. 2017...is high, it’s generally not as long lasting as AAV (up to several weeks vs. AAV’s several years). But these... To add insult to injury, retroviruses generally have moderate to high immunogenicity, meaning they induce...interchangeable between the two systems, other, more general, viral components such as a heterologous envelope...AAV has many excellent features for human gene therapy and is currently being used in clinical settings...adenovirus have been identified so far! Adenovirus generally has high immunogenicity, but some strains have... fact: All viruses discussed in this blog are generally broadly infective, with one exception. Blood cells...

... 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...