We narrowed to 109 results for: 1-Oct
-
TypeBlog Post...) element and the cis-acting EBNA-1 from Epstein-Barr virus. EBNA-1 binds oriP and allows for replication...Difficult to purify reprogramming proteins. Table 1: Key Features of Different Methods for Generating ...-based expression requires serial transfection of 1 or 2 plasmids that express the reprogramming factors... Replicating oriP/Epstein-Barr nuclear antigen-1 (EBNA1) based episomes: These plasmids carry the origin...method Most iPSC research falls into two categories: 1) studies focused on better understanding the mechanisms...iPSCs experiences in the comments below! References 1. Sano, Masayuki, Minoru Iijima, Manami Ohtaka, and...Methods figures were adapted from this reference. 1. Malik, Nasir, and Mahendra S. Rao. "A Review of the...
-
Optimizing Donor DNA for Enhanced CRISPR Genome Editing
TypeBlog Post... Figure 1 below) was free to anneal to complementary ssDNA molecules in vitro. Figure 1: Cas9 binding...allowed sequence replacement rates of approximately 1%. This is by no means a large number, but it was attained...who worked on this publication: References 1. Richardson, Christopher D., et al. "Enhancing homology-directed...contributed by guest blogger Chris Richardson, a Postdoctoral Researcher in Jacob Corn’s lab. CRISPR-Cas9 ... optional donor DNA. Chris Richardson is a Postdoctoral Researcher in Jacob Corn’s lab investigating... -
28 Hot Plasmid Technologies from 2015
TypeBlog Post...your target gene! Tanenbaum et al., Cell. 2014 Oct 8. pii: S0092-8674(14)01227-6. Multi-bit genetic...light-sensitive LOV2 domain of Avena Sativa phototropin 1 (AsLOV2). In the dark, this peptide is caged by the...iLID nano and 800nM to 47µM for iLID micro; Figure 1). With this great affinity range, these LIDs have ...Figure 2). They are all available now at Addgene. 1 Lungu et al., Chem Biol. 2012 Apr 20;19(4):507-17....Guntas et al., Proc Natl Acad Sci USA 2015 Jan 6; 112(1):112-7. doi: 10.1073/pnas. CRISPR-Cas9 optogenetic... al., BMC Molecular Biology. 2013 August 20th; 14(1). TRICK: A method for visualizing the first round...for a variety of experimental purposes including (1) single or multiple, constitutive or inducible, gene... -
Starter guide to induced pluripotent stem cells (iPSCs) part 1: A renaissance in regenerative medicine
TypeBlog Post...into more specialized cells with specific functions [1]. In many tissues, stem cells serve to replenish/replace...damaged cells that no longer function adequately [1]. Stem cells’ ability to differentiate into multiple...://www.linkedin.com/in/kmukherjeephd/. References 1. Dittrich, R., M.W. Beckmann, and W. Wurfel, Non-embryo-destructive...The potential of muscle stem cells. Dev Cell, 2001. 1(3): p. 333-42. PubMed PMID: 11702945. 6. Cowan, C....preimplantation genetic diagnosis. Hum Reprod, 2008. 23(1): p. 46-53. PubMed PMID: 17989069. 8. Takahashi, K... the ectopic expression of four embryonic genes- OCT4, SOX2, KLF4, and C-MYC. iPSCs were first generated...communication and publishing after completing her postdoctoral training from Massachusetts General Hospital... -
Using Phosphoserine to Study Protein Phosphorylation
TypeBlog Post...cellular proteins are modified through phosphorylation (1). However, the expansion of studies on protein phosphorylation...three components required for translation (Figure 1): A tRNA that recognizes a specific ‘open’ codon ... open reading frame (ORF) of interest (Figure 2). 1. The phosphoserine OTS The backbone of the phosphoserine...modification allowed the deletion of release factor 1, the protein responsible for terminating translation...Rinehart lab: pCRT7 NT TOPO E17TAG GFP and pGEX-6P-1. The pCRT7 NT TOPO vector contains a mutant pSer-containing... can follow her on Twitter @nieminm. References 1. Cohen, Philip. "The origins of protein phosphorylation...contributed by guest blogger Natalie Niemi, a postdoctoral fellow at the Morgridge Institute for Research... -
Starter Guide to induced Pluripotent Stem Cells (iPSCs) Part 2: Reprogramming and Transdifferentiation
TypeBlog Post...type of choice in vitro is known as reprogramming [1]. The process can be divided into two stages: Dedifferentiation...//www.linkedin.com/in/kmukherjeephd/. References 1. Hochedlinger, K. and R. Jaenisch, Nuclear reprogramming...from human peripheral blood. Cell Stem Cell, 2010. 7(1): p. 15-9. PubMed PMID: 20621044. PubMed Central PMCID...induced pluripotent stem cells. Hepatology, 2010. 51(1): p. 297-305. PubMed PMID: 19998274. PubMed Central... pluripotent stem cells. Nat Biotechnol, 2014. 32(1): p. 84-91. PubMed PMID: 24291815. PubMed Central ...keratinocyte lineage. Methods Mol Biol, 2014. 1195: p. 1-12. PubMed PMID: 24510784. PubMed Central PMCID: PMC4096605...induced pluripotent stem cells. PLoS One, 2010. 5(1): p. e8763. PubMed PMID: 20098701. 31. Tang, Z.H.,... -
Cancer and the Immune System: Deciphering the Relationship
TypeBlog Post...such as IL-8, IFNγ, and colony stimulating factor 1 (CSF-1). Additionally, tumor cells recruit regulatory...introduced the “Three Es of Immunoediting” nomenclature. 1. Eliminate - Latent tumors are recognized by effector...the tumor, activates hypoxia inducing factor (HIF-1), which promotes the production of VEGF to increase...This study used CRISPR gene editing to knockout PD-1 on immune cells harvested from patients and thereby...exploring new avenues in healthcare. References 1. Vinay, Dass S., et al. "Immune evasion in cancer:...lymphoblastic leukaemia in children and young adults: a phase 1 dose-escalation trial." The Lancet 385.9967 (2015)...contributed by guest blogger Subhadra Jayaraman, a doctoral candidate at Binghamton University Cancer is one... -
Designing Your Chalk Talk for the Academic Job Interview
TypeBlog Post...acceptable. Aims should be mechanistic, doable by 1-3 people in 1-5 years, and contribute to resolving the overarching...possible, do at least two full length practices (~1 h) and get feedback. It’s also helpful to practice...contributed by Erik Snapp, the Director of Student and Postdoctoral Programs at the Janelia Research Campus of the...from Oregon Health Sciences University, did his postdoctoral fellowship with Jennifer Lippincott-Schwartz...and now serves as the Director of Student and Postdoctoral Programs at the Janelia Research Campus of the... -
Avoiding the Dark Side of Fluorescent Protein Fusions with mOX FPs
TypeBlog Post...experiments or even negatively impact cells (see Figure 1). When behaving properly, FPs enable researchers ... disulfide bonds in the secretory pathway (Figure 1) [4,6–8]. Additionally, FPs that encode N-glycosylation... sarcoma-associated herpesvirus. References 1. Snapp, Erik. "Design and use of fluorescent fusion...from Oregon Health Sciences University, did his postdoctoral fellowship with Jennifer Lippincott-Schwartz... from Albert Einstein College of Medicine. Her doctoral research was conducted in the laboratory of Erik...subcellular environment. Currently, she is a SPIRE postdoctoral fellow at the University of North Carolina at... -
X-CHIME: Context Dependent Germline Knockout in Immune Cells
TypeBlog Post...in the immune system. Nature Immunology, 25(1), Article 1. https://doi.org/10.1038/s41590-023-01689-6 ... strains and lentiviral expression vectors (Table 1). The four systems Name Function Plasmid ... GFP (vex) Rosa26-FlpO-ERT2; H11-Cas9 Table 1: X-CHIME systems Proof of Concept These systems were...was lethal (LaFleur et al., 2024). Figure 1: Schematic showing the X-CHIME approach. Figure from...genes in the immune system. Nature Communications, 10(1), 1668. https://doi.org/10.1038/s41467-019-09656-2... X-CHIME plasmids here! Marty LaFleur is a Postdoctoral Fellow in Arlene Sharpe’s laboratory at Harvard... -
New Tools Enable CRISPRa for Neuroscience Applications
TypeBlog Post...by mating with Cre-expressing mice. Figure 1: (A) Schematic of the Cre-dependent SPH construct ... alpha (aCaMKII; excitatory neurons) and Synapsin-1 (pan-neuronal) promoters (Figure 1b), and sgRNAs can...ubiquitous expression of sgRNA(s) and mCherry (vector 1) with neuron-specific expression of dCas9-VPR (Figure...Neuron-specific expression is mediated by the synapsin 1 (SYN) promoter, which was superior in its ability ...Patrick Miller-Rhodes, a Ruth L. Kirschstein NRSA Predoctoral Fellow at University of Rochester Medical Center...Miller-Rhodes is currently a Ruth L. Kirschstein NRSA Predoctoral Fellow at University of Rochester Medical Center... -
Screening for Successful Genome Editing with Digital PCR
TypeBlog Post...digital) signal at the end of the reaction (Figure 1). Partitions that did not contain any target DNA at...integrated through homology-directed repair (HDR) (1), or indel mutations resulting from non-homologous...Follow him on Twitter @AnotherLabRat. References 1. Miyaoka, Y. et al. Isolation of single-base genome-edited...This post was contributed by Scott Findlay, a Postdoctoral Fellow at the University of Alberta. If you’... out of frame! Scott Findlay is currently a Postdoctoral Fellow at the University of Alberta. He is interested... -
Tips for Getting a Faculty Position
TypeBlog Post...Full "Applying for a Faculty Position" eBook Here 1. Have a competitive CV You don't necessarily need ...that people would agree is important to understand. (1-2 sentences). For example: Distinguishing correctly... What will be the focus of your first grant? (1-2 sentences) Simply stated, what are your aims... potential collaborations, critiques of your postdoctoral research, etc. The second question is whether...from Oregon Health Sciences University, did his postdoctoral fellowship with Jennifer Lippincott-Schwartz...and now serves as the Director of Student and Postdoctoral Programs at the Janelia Research Campus of the... -
Supporting Rare Disease Research with BeHeard 2017
TypeBlog Post...estimated to affect anywhere from 1 in 6,000 to 1 in 19,000 live births (1). While disruption of the smn1... funding opportunities. References 1. Jones, C., et al. "PP09. 1–2352: Systematic review of incidence...Peritoneal Mesothelioma Noushin Nabavi is a Postdoctoral Fellow at the Vancouver Prostate Center and ... -
Changing Labor Laws Bring Increased Postdoc Wages
TypeBlog Post...$47,476 and the date for implementation was set for Dec 1 2016. In order to be exempted from the need to pay... these institutions (populations potentially from 1 to 5761, according to the NSF data) and therefore ...exactly how postdoctoral salaries are changing. Transparent salary information for postdoctoral positions...institutions set their postdoctoral salary scales to the NRSA scale (Figure 21, National Postdoctoral Association...Stanford raised salary levels to $50,000, and as of October 1st the salary minimum was raised again to $51,...as having postdocs, representing ~85% of the postdoctoral workforce. Based on our communications with ...according to the NSF data, account for 18% of the postdoctoral workforce in the sciences. The Dana Farber Cancer... -
Cancer, Inflammation and Immunity - Harnessing the Body’s Defenses to Fight Cancer
TypeBlog Post...aggregation, and potentiates the effect of PD-1 blockade.“ (Ref 4). PD-1 helps tumor cells evade the immune system...fasting has a positive effect on cancer treatment (Ref 1). As described in the presentation, starvation or ...of the immunosuppressive cell surface receptor PD-1. This immunosuppressive environment in turn may represent... (Chimeric Antigen Receptors) CAR T cells and PD-1 modulators. An exciting development in immunotherapy...free deposit and distribution service! References: 1. Pietrocola, Federico, et al. "Caloric restriction...Relationship” by our guest blogger Subhadra Jayaraman, a doctoral candidate at Binghamton University! Cancer therapeutics... -
Harnessing Bacterial Toxins for Allelic Exchange
TypeBlog Post... requires selection. These two steps are (Figure 1): The initial plasmid integration step (the so-called...antibiotic resistance marker from the vector (abR, Figure 1). The resolution step (“double crossover”) which leads... loss of a specific phenotype (using sacB, Figure 1, further discussed below). Counter-selectable marker...Current protocols in molecular biology 106.1 (2014): 1-16. PubMed PMID: 24733238. Additional resources on...This post was contributed by Jacob Lazarus, a postdoctoral researcher at Harvard. There’s an astounding...is now an attending physician. He performs post-doctoral research in the lab of Matthew Waldor at Harvard... -
When is a Monomer not a Monomer? The Top Three Ways Your Favorite Fluorescent Protein Oligomerizes in Cells
TypeBlog Post...Figure 1). Figure 1: Each problem with dimerizing fluorescent proteins is illustrated. 1) Membrane... in nature are prone to dimerization (i.e. EGFP) [1, 2] or even form obligate tetramers (i.e. DsRed) [...active state due to closure by FP dimerization. 1. Transmembrane fluorescent protein fusions Transmembrane...bright, non-physiological, and difficult to ignore [1, 2]. 2. Fusion to an obligate dimer or oligomer Several... For a more detailed discussion see [5]. Problems 1 and 2 depend on protein concentrations. An imperfect... distance runner, gardner, and cook. References 1. Snapp, E.L., et al., Formation of stacked ER cisternae...from Oregon Health Sciences University, did his postdoctoral fellowship with Jennifer Lippincott-Schwartz... -
The Future of Research Symposium Boston 2015
TypeBlog Post...all of its successes, has many deep-rooted flaws (1). As we discovered through the four workshops at the... Follow David on Twitter @driglar. References 1. Alberts, Bruce, et al. "Rescuing US biomedical research...the Future of Research will be held from 22-24 October. This blog has been contributed by guest blogger...strong supporter of increasing the number of postdoctoral scientists at University of Maryland. Paula... can register here or live stream the event on October 22nd and 23rd here. Many thanks to our guest blogger... -
Scientific Peer-review: Providing Critical and Kind Feedback and Advocating for Open Science
TypeBlog Post...the journal-specific guidelines. Peer-review step 1: The first read At the first read of the paper, I ...group of Prof. Mark Tester, where she focuses on (1) salt-induced changes in root-to-shoot ratio in Arabidopsis...post was contributed by Magdalena Julkowska, a postdoctoral researcher at KAUST, Saudi Arabia. From the... submission to the subsequent journal. As a postdoctoral researcher studying plant development and abiotic...