...Koeris' own transition from graduate student to entrepreneur, listen to our full interview with Dr. Koeris...contrasts being a graduate student with being an entrepreneur 29:00 - 32:20 : What did It mean for Dr. Koeris...developing a diagnostic? 36:53 - 38:00 : As an entrepreneur, is it more important to develop something that...Board and how he hopes to help young biotech entrepreneurs.   We'd like to thank Dr. Koeris for taking... entrepreneur...

...school help prepare you for the roles and skills necessary to be a succesful entrepreneur? As a co-founder...explained that the data must be reproducible by someone else. Second, "an entrepreneur must be OK with the constant...its pitch the best, wins. He explained that entrepreneurs must clearly communicate their idea at various...constantly practicing. “The expenditure of effort in preparation and practice is directly related to success presenting...from the early 1990s that were free and free to repair.” If the team works well, the idea is solid, and...

...performed using the transfer plasmid, a plasmid encoding Rep and serotype-specific Cap genes, and a plasmid encoding...order to generate high quality preparations. After production, all virus preps are titered and subjected to... containing) AAV particles within representative vector preparations was quantified with electron microscopy...typically prepared using endotoxin-free plasmid purification kits. Resources Browse lentiviral preps available...encoding adenoviral helper sequences. AAV preparations are purified by iodixanol gradient ultracentrifugation...purity and titers adequate for in vivo studies. Preparations are then aliquoted and stored at -80 °C. Titer...on viral genomes isolated from the final AAV preparation. Sequencing results are analyzed to confirm the...

...modulate iPSC reprogramming efficiency. Listen to the segment on modulating iPSC reprogramming  Hot Plasmid...Out All of Our Hot Plasmids Articles Browse the Repository  ...

...promote bone regeneration in 3D biomaterials. Sci Rep, 2016. 6: p. 26761. PubMed PMID: 20065292. PubMed...cells to mature functional Purkinje neurons. Sci Rep, 2015. 5: p. 9232. PubMed PMID: 25782665. PubMed ...that have been reprogrammed to iPSCs successfully [10-12]. The next stage of reprogramming consists of redifferentiation...-42]. Reprogramming and transdifferentiation: When to use one over the other Both reprogramming and transdifferentiation...processes by which cells are converted to replenish and repair cells that are lost or damaged. Both transdifferentiation...transdifferentiation, and reprogramming: future directions in regenerative medicine. Semin Reprod Med, 2013. 31(1...goal in the field of regenerative medicine is to replace lost or damaged cells. Here, I will discuss the...

...Problems with FRET kinase reporters Difficult to Multiplex: FRET reporters are comprised of two fluorescent...phosphatase-accessible. Kinase Translocation Reporters (KTRs) Kinase Translocation Reporters (KTRs) are genetically encoded...single-cell kinase activity is FRET, but FRET reporters are challenging to design and difficult to multiplex...alternative tool with their Kinase Translocation Reporters (KTRs) whose cellular localization serves as a...multiplexed together. Challenging  to Design: FRET reporter activity depends on both the distance and the .... Slower Dephosphorylation Rates: FRET-based reporters can have slower dephosphorylation rates than KTRs...KTRs which is hypothesized to be caused by FRET reporters having phosphorylated conformations that are less...

... when Yamanaka and colleagues reported that it is possible to reprogram differentiated cells into induced...starting cells for reprogramming while PBMCs collected from blood are reprogrammed immediately after collection...modified reprogramming factor cocktail, and viral protein EBNA-1. Methods for reprogramming hiPSCs from...the common goal of all reprogramming methods is the expression of the reprogramming factors- OCT3/4, SOX2...long-term expression of the reprogramming factors. However, reprogramming of PBMCs by episomal vectors...source of somatic cells for reprogramming. Enhancing PBMC reprogramming efficiency: the pCXLE episomal ...of an shRNA for P53 and replacing C-MYC with L-MYC in the episomal reprogramming cocktail by Okita et al...

...controllable and reproducible set up,” Bernach explains. Developing an artificial leaf replica means creating... the TIMER growth rate bioreporter. During protein maturation, the bioreporter spontaneously changes from...sensor with proof-of-principle experiments that replicate current knowledge, but the sensor can also kinetically...  Michal Bernach: Developing an artificial leaf replica system to study plant-bacteria and bacteria-bacteria..., Bernach sought to develop an artificial leaf replica system. “This bottom-up approach will allow me ..., Bernach plans to use a suite of fluorescent reporters. Bacteria containing unstable fluorescent proteins...artificial leaf, Bernach will use the CUSPER bioreporter such that fluorescent proteins expression is ...

...identification of a reagent is crucial for reproducibility because mistakes in this can lead to wasted...Object Identifier (DOI). Nearly all the papers or preprints you read or produce have one. The DOI serves as...these metadata and publications help to enable reproducibility of experiments. Additionally, proper citation...address. For the RRID format, lenti-Cas9-VQR is represented as “RRID:Addgene_87155” and the following URL... the journal’s specific author guidelines for reporting your materials. If you deposit a plasmid with ...to ensure that materials available through our repository are verified and identifiable to save researchers...researchers time and make their work more reproducible. While Addgene plasmid URLs act as unique PIDs, we’re...

...a library of reporter plasmids, each containing two fluorescent proteins and four repeats of a miRNA target..., it will repress expression of mKate2 relative to EBFP2. Weiss’s group used these reporters to define...collection of reporter plasmids is amenable to Golden Gate assembly for turning the single-input reporters into...highlight a subset of the new plasmids in the repository through our hot plasmids articles. These articles...Neuron. 2019. PubMed PMID: 30704911. Acoustic reporter genes for noninvasive imaging of microbes in mammalian...contributed by Lukas Morgan  Listen to the acoustic reporter gene podcast segment In recent years, scientists...heavily on fluorescent protein genes. These optical reporters have limited deep-tissue performance, and sometimes...

...Find plasmids expressing tTALE-FP Shin et al., Sci. Rep. 2019. https://doi.org/10.1038/s41598-019-53722-0...transcriptional reporter consisting of a quickly-degraded destabilized GFP variant fused to a 4X CDRE repeat. In ...highlight a subset of the new plasmids in the repository through our hot plasmids articles. These articles...infection, because this setup allows for experimental replicates for each of the gRNAs. The idea was to assign...remove a stop cassette upstream of an effector/reporter gene. In cells where the stop cassette is removed...organisms including Drosophila, fish, and mice, by replacing the effector with a gene of interest in the cells...https://doi.org/10.1101/788679 A destabilized GFP reporter for calcineurin activity in yeast Article contributed...

... the Doyon Lab’s publication: Dalvai et al. Cell Rep. 2015 . Alternatively, cDNAs can be cloned directly...Zebrafish Xenopus CRISPR Resources CRISPR Guide Viral Preps Protocols gRNA Design Tools CRISPR Blog Posts The... CRISPR 101 post for more information about HDR/repair template design. How to use CRISPR to tag your ...plasmids may be used. The HDR (homology directed repair) donor plasmid with homology arms and selection...transcription factors with FLAG in human cell lines. To repeat the tagging in your system, use the plasmids as...introduces an N- or C- terminal affinity tag (3xFLAG-2xSTREP) on endogenous genes for the isolation of native...the AAVS1 locus. Doyon Tagging Plasmid: 3xFLAG-2xSTREP Construct for integrating at the AAVS1 "safe harbor...

...packaging proteins for AAV are rep and cap. Rep is required for viral replication and also aids in infrequent...release from late endosomes when AAV is trafficked. Rep and cap mediate AAV pseudotyping which allows for...ITRs serve as a type of origin of replication to initiate replication of the dsDNA virus. Viral packaging...plasmid while the other plasmids with packaging or replication components are called helper plasmids. Below,...this promoter-gene combo are two long terminal repeats (LTRs). These LTRs define the limit of what will...different in sequence. Both LTRs do share common repeat regions in addition to their unique regions as ...constructs. Sandwiched between two inverted terminal repeats (ITRs) is the promoter and cargo, in that order...

...Mutations Both Inhibit HIV-1 Replication and Accelerate Viral Escape. Cell Rep. 2016 Apr 5. PubMed PMID: ...reactivation of the HIV-1 latent reservoirs. Sci Rep. 2015 Nov 5;5:16277. PubMed PMID: 26538064. PubMed...-lymphoid Cells by CRISPR/Cas9 Gene Editing. Sci Rep. 2016 Mar 4;6:22555. PubMed PMID: 26939770. PubMed...DNA efficiently suppresses hepatitis B virus. Sci Rep. 2015 Jun 2;5:10833. PubMed PMID: 26035283. PubMed...designed multiple gRNAs to target the 5’ long terminal repeat (LTR) of the HIV-1 genome, which acts as a promoter...create mutations in genes essential to HIV-1 replication. However, this approach could permit some viral...for CRISPR HIV-1 therapy Both of these methods represent exciting advances in preclinical HIV-1 research...

...protein tracking in vivo. Sci Rep. 2015 Jul 7;5:11771. doi: 10.1038/srep11771. PubMed. McKinney SA, Murphy...cytoplasmic strands. Image from Sattarzadeh et al., Sci Rep. 2015 CC BY 4.0 Applications and limitations of GFP...conduct subcellular localization studies or make a reporter construct. Fluorescent proteins (FPs) are also...and it would be expensive and time-consuming to repeat the process with these new photoconvertible proteins...culture with cytosol-targeted GFP. White circles represent the irradiated area. Photoconverted GFP spreads...investigators when using forms of mEos." The ability to repurpose GFP is great news for microscopy enthusiasts! ...

...Reveals Local and Long-Range Circuit Motifs. Cell Rep. 28(13):3450-3461. PubMed (Link opens in a new window...novel population of long-range inhibitory neurons. Preprint (Link opens in a new window) Yu K, Ahrens S, Zhang...

...Murakoshi H, et al. Sci Rep. 2015. Pubmed PMID: 26469148. Murakoshi H, et al. Sci Rep. 2017. Pubmed PMID:...highlight a subset of the new plasmids in the repository through our hot plasmids articles. These articles...microbes, used coffee grounds could be economically repurposed as starting material for the bioproduction of...

... to produce AAV: 1) the RepCap plasmid which provides the AAV replication (rep) and capsid (cap) genes...genes. AAV replication uses the host’s polymerase, but requires Rep proteins to process a double-stranded...adenovirus genes which help mediate AAV viral replication; and 3) the transfer plasmid which encodes a ...still necessary. To save time and money, I like to miniprep 5 mL of my AAV transfer plasmid cultures and then...pellet for the remainder of the cultures and later maxiprep only the cultures with intact ITRs. I use a spreadsheet...culture supernatant because it’s time consuming to prepare the 40% PEG solution. PEG takes several hours on.... Rather than investing all of this effort in preparing reagents, I choose to only harvest the cell pellet...

...Chayama K, Yamamoto T. Sci Rep. 2014 Jun 23;4:5400. doi: 10.1038/srep05400. PMID: 24954249 Find the Mulitplex...University who described their new tool in Scientific Reports in June. “The multiplexity is one of the most advantageous...will also allow simultaneous activation and/or repression of multiple genes, and more. The researchers ...

...Mapping Multiprotein Complexes in Human Cells. Cell Rep . 13(3):621-33. PMID: 26456817 Fu Y, Sander JD, Reyon...other hand, if a repair template with homology arms was supplied, the break could be repaired according to...loci. Activate or Repress : dCas9 fused to transcriptional activators or repressors can modulate transcription...mouse, and rat) Bacteria ( E. coli , Streptococcus , Streptomyces , and others) Drosophila Plants (monocots...Clustered Regularly Interspaced Short Palindromic Repeat) sequences were initially discovered in the E. ...Protospacers are separated by short palindromic repeat sequences. (1) The CRISPR array is transcribed ...tracrRNA) with homology to the short palindromic repeat. The tracrRNA helps recruit RNAse III and Cas9 ...