We narrowed to 814 results for: Ran;

...errors like uneven pipetting or irregular membrane transfer. Figure 1: A loading control is a type...lies the linear detection range. Figure 4: The linear detection range of the assay is between the.... No pipetter is perfect, and proteins may not transfer precisely evenly across the entire blot. So how...directly correlate to protein abundance on your membrane, which would be directly reported by band intensity... in the loading control are due to loading and transfer errors — not to any factors from before the proteins...control points are outside of the linear detection range. As a result, they have almost the same value on...computational quantification. Outside the linear detection range, band intensity measurements become extremely susceptible...

...quantify RNA transcription by RNA Polymerase III. By expressing Corn under promoters for transcripts uniquely... and orange fluorescent aptamers Corn was described in the same study as Red Broccoli and Orange Broccoli...fluoresce in the red and orange channels, respectively. Both Red Broccoli and Orange Broccoli are derived ...multiple derivatives of thiazole orange (TO) and fluoresces in the orange or red channel, depending on the...aptamers are used to track cellular processes by transforming cells with plasmids encoding aptamer-tagged ...tagging RNA sequences with an aptamer and RNA transcription by placing the aptamer downstream of a known...hydroxybenzylidene imidazolinone (DFHBI). DFHBI is membrane permeable and non toxic to cells. Because it rapidly...

...new class of sensors for neuropeptides and neurotransmitters. Motivated to develop a biosensor for oxytocin...devised a “Modular fluorescence unit fused with TRansmembrane region-to-IntrAcellular loop linkers” (MTRIA...sensors based on GPCRs recognizing various neurotransmitters, neuropeptides and other ligands - almost ...kinetics and localization for imaging synaptic transmission. bioRxiv 2022.02.13.480251. doi: 10.1101/2022.02.13.480251...2022.02.13.480251 JEDI-2P voltage sensor The François St-Pierre lab and colleagues have developed a new...they observe fluorescence localizes to the plasma membrane only when the RTK is activated and the ITAM phosphorylated...relies on a change in localization to the cell membrane and not a direct change in the fluorescent properties...

...permeabilize the cell membrane to allow antibodies to pass through the cell membrane. Formaldehyde fixation...dye emits green, yellow and orange photons while PE emits yellow and orange photons. If FITC and PE were...sources that allow users to assess a cell’s size, granularity, and target protein composition. It is used for...target protein expression levels, assessing post-translational modifications, determining cell health, analyzing...a pressurized buffer called sheath fluid and transported through tubes or capillaries to a laser. As the...degree of internal complexity such as extensive membranes produces a greater SSC.  Protein expression, measured...targets, measured by fixing and permeabilizing membranes For intracellular protein targets, users must ...

...lateral”) gene transfer. Horizontal gene transfer can occur naturally through transformation (cellular uptake...? Plasmids 101: Transformation, Transduction, Bacterial Conjugation, and Transfection Resources on Addgene.org...environment), transduction (viral infection by bacteriophages), or conjugation (direct transfer from one cell...one popular method scientists use to promote transformation, but it isn’t just for lab specimens — there... evidence that lightning can also promote gene transfer in soil bacteria! (Demanèche et al., 2001) Beneficial...humans only noticed last century! Nevertheless, the transfer of plasmids carrying these genes between bacteria...plasmid are now used routinely in biotechnology to transform plant cells. Degradative plasmids: microbial hardiness...

...mouse cell type specific transcription factor binding sites interspaced with random sequences.   Strategy...synthetic promoters. Legend: TSS: transcription start site. TFBS: transcription factor binding site. Image from...particular cell type is important not only for translational research, such as gene therapy, but also basic... using naturally occurring promoters to drive transgene expression, the Roska lab built synthetic promoters...conserved sequences that are upstream of the transcription initiation sites of at least two genes with ...designed using stretches of DNA that were transcriptionally active in retinal cell types in mice. ...synthetic promoter driving expression of the same transgene: an optogenetic reporter, channelrhodopsin, fused...

...produced at high titers, can accommodate large transgenes, transduce quiescent and dividing cells, and do not..., which results in the death of transduced cells and loss of transgene expression (Interestingly, the ...adenoviral genome. The genome itself is a linear double-stranded DNA of ~36kb, with inverted terminal repeats (...E3 regions and can accommodate up to 8.2 kb of transgene DNA. The second generation vectors are deficient...cloning capacity is limited to 8.2 kb, and in vivo transgene expression ceases relatively quickly due to immune... the E1 gene from the packaging cell line is transferred into the AdV by recombination. It is therefore...adenovirus is required to provide the viral proteins in trans. The Gutless AdV have generated high interest for...

...Tethering dCas9 to the histone acetyltransferase p300 or the DNA methyltransferase DNMT3A allows researchers...affect transcription independent of their enzymatic activity. They interact with transcriptional machinery...on DNA methyltransferase 1 (DNMT1) to copy methylation marks from parental to daughter strands of DNA....specifically edit methylation sites.  Step aside, transcriptionally active dioxygenase  When I started to work...by distinct proteins and result in unclear transcriptional consequences (Sprujit 2013). Enzymatic domains...in the genome and directly interacting with transcription machinery.   Well, I’ve got some good news...rounds of cell division, the original methylated strand is diluted to undetectable amounts: ergo, DNA demethylation...

...BioRender.com. Transformation Grade DNA (low quality/low quantity) – Bacterial transformation protocols require...highest-competency cells you will still struggle to get transformants if the plasmid you are using is completely degraded...important reactions like PCR or restriction digests. Transfection Grade DNA (high quality/high quantity) – Depending...Depending on the experiment, mammalian cell transfections can require quite large amounts of DNA (easily...carry-over into the eluate, while other kits are guaranteed to yield endotoxin-free DNA. Once you know the...kits are designed to accommodate specific volume ranges of culture as their input (Table 1). When you feel...addition to standard purification kits, there’s a range of specialty kits available for plasmid purification...

...and membrane integrity (Fig. 2). For example, saponin interacts with cholesterol in cell membranes and...can lose membrane-associated proteins, but it is better at permeabilizing organellar membranes. Created...red lines), which maintains structural epitopes (orange region), but it can block epitopes hidden within...proteins that need a little help crossing cell membranes as they are not able to diffuse across natively...Permeabilization essentially punches holes in the cell membranes of your sample to allow antibodies better access...target is already on the extracellular side of the membrane, then this step may not be necessary. In fact,...and leaves many membrane-associated proteins in place. In contrast, Triton X-100 and Tween-20 are examples...

...prevent efficient transcription from sgRNA targets. CRISPRi is effective in a narrow range around the transcription...two sets of sgRNAs; A, the top 3 ranked sgRNAs, and B, the next 3 ranked sgRNAs. Dolcetto was able to discriminate...Cas9 derivatives. Unmodified Cas9 causes double stranded DNA breaks that commonly result in a complete ... lab selected sgRNAs in this optimal window and ranked them based on their optimized sgRNA rules and potential...gene expression, dCas9 can also be used for transcriptional activation. Activation of specific genes allows... strategies have been developed to recruit transcriptional machinery to dCas9 and its RNA directed binding...include RNA domains that recruit additional transcription factors. The Root and Doench labs utilized tracrRNAs...

...Biological Material Transfer Agreement (UBMTA) is one of the most commonly used ones for transfers between academic...for more information. Happy transferring!    Many thanks to our Tech Transfer team members Maura Coughlin...Addgene, then you’ve been involved in the material transfer agreement (MTA) process. Even though this process...curious about the hows and whys of the material transfer agreement, read on!  What is the MTA?  An MTA...the creation of a standardized agreement on the transfer of biological materials in 1995: the UBMTA. Because...most academic and nonprofit institutes, the Tech Transfer department is responsible for managing research...plasmid with Addgene, your organization’s tech transfer office is sent a deposit agreement. The One-Time...

...also incorporated LOV-Turbo into TransitID experiments.  Find TransitID plasmids here! These two developments...genetic cargo Optogenetic proximity labeling and TransitID Tracking cellular history with protein ticker ...in vitro and in vivo. Incorporating reverse transcriptase (RT) variants, PE6a and PE6b are 500–800 bp ...be tethered on the viral surface, expanding the range of targetable cell types. DIRECTED is compatible...37612276.   Optogenetic proximity labeling and TransitID By Mike Lacy Proximity labeling can identify protein...plasmids here! Next, Qin and Cheah et al. described TransitID, where two proximity labeling enzymes, TurboID...catalyzing the biotinylation of nearby proteins. B) TransitID uses sequential labeling by TurboID and APEX2 ...

...of Pennsylvania Vector Core. Transfections are performed using the transfer plasmid, a plasmid encoding...PCR with primers targeting the transfer plasmid used in the transfection. PCR products are analyzed by ...cell cultures are inspected for sterility. Transducibility Some viral vectors are tested in vitro and ...images).     Figure 2: AAV Pro cells were transduced with either pAAV-Ef1a-mCherry-IRES-Cre (55634...filtration through a 0.45 µm polyethersulfone (PES) membrane. The preparations are then aliquoted, frozen, ...from Wang et al., 2018). Briefly, 293T cells are transduced with serial dilutions of a lentiviral vector,...taken offline and discarded. Confirmation of the transfer plasmid Addgene uses a rigorous barcode matching...

...fused to the N-terminus of LSSmOrange pLSSmOrange-C1 Orange Mammalian Express a gene of interest fused...GFP variant) commonly used with mRuby2 pLSSmOrange-N1 Orange Mammalian Express a gene of interest fused...Bioluminescence Resonance Energy Transfer CRET= Chemiluminescence Resonance Energy Transfer    Interesting Articles...FRET, aka Fluorescence (Förster) Resonance Energy Transfer, technique which allows the detection of molecules...distance-dependent physical process by which energy is transferred non-radiatively from an excited molecular fluorophore... (the acceptor) by means of intermolecular long-range dipole-dipole coupling." The emission of the acceptor...fused to the C-terminus of LSSmOrange mRuby2 Red Mammalian Expresses mRuby2 (a RFP variant) commonly used...

...flow of electric charges across cell membranes and maintain membrane potential in response to visible light...across the axon's membrane: once a certain number of positive ions crosses the cell membrane, a threshold ...rhodopsins enable ions to flow across the axon’s membrane, thereby controlling neural activity (Bernstein...the opsin into the correct orientation to be transcribed (Figure 2) (Sharma and Zhu, 2014). How does FLEx... optogenetic FLEx vectors in combination with transgenic animals or rAAVs expressing Cre under a specific...the mechanisms of sleep and anesthesia. In Cre transgenic mice, they targeted the dopamine (DA) neurons... vectors combined with the use of several Cre transgenic mice targeting different brain areas enabled ...

... technologies have transformed the palette of experiments possible in a wide range of organisms and cell...genomic locus (CRISPR-ChAP-MS) could further transform the range of experiments imaginable in C. elegans. ...Genome Editing with Short Single-Stranded and Long, Partially Single-Stranded DNA Donors in Caenorhabditis...at 25 ºC. This method resulted in efficiencies ranging from 11-100% of F1 animals carrying precise knock-ins...This stringent selection allowed me to optimize a range of editing parameters: oligo repair templates with...with homology arms of 35-80 bp, and DNA double-strand breaks up to 54 bp from the desired insertion site..., they devised guidelines for designing single-stranded repair templates (Farboud et al., 2019).   The...

... pair has focused considerable attention on transcription factors, starting with an established method... relies on antibodies to recognize and bind transcription factors directly, but that’s a slow-going process...ENCODE project is to look at all 1,500 or 1,600 transcription factors and figure out where they bind to the... ChIP-seq, but it’s really hard for lots of transcription factors. Our groups got together to adapt CRISPR...big difference. We can now look at many more transcription factors. Addgene: How does the system work? ...second plasmid to target an epitope tag onto the transcription factors. By the way, you can actually use this... in all kinds of different assays, not just transcription factors. The second vector is a homology directed...

...A double-strand DNA break occurs within the core spacer of each loxP site and the two strands are ligated...maintained).  Translocation: If the sites are on separate DNA molecules, a translocation event is generated... expression – placing a series of transcriptional and translational termination features with loxP sites... pun) specific, targeted DNA modifications in transgenic animals, embryonic stem cells, and/or tissue-...that there is virtually no chance of it occurring randomly. Therefore, inserting loxP sites at deliberate...Inversion: If the loxP sites are on the same DNA strand and are in opposite orientations, recombination...research tool, especially in the field of mouse transgenics. A few of the most common uses are listed below...

...which ensures the editor will only be transiently present in transduced cells, limiting off-target editing... dCas9 fused to a DNA methyltransferase — that can drive lasting transcriptional silencing (Nuñez et al...virus-like particles (eVLPs) offers an option to transduce cells directly with ribonucleoprotein cargo. eVLPs...attB/P recombinase landing sites on opposite DNA strands. When the prime editor releases the DNA, the two.... (2024). Engineered virus-like particles for transient delivery of prime editor ribonucleoprotein complexes...Histone tail for Autoinhibition Release of Methyltransferase (CHARM), with several advantages over the ...  Rather than overexpressing an exogenous methyltransferase for epigenetic silencing, which can be toxic...

...et al., 2018). Unbiased, random insertions can be generated with a transposon-based strategy (Sheridan...fluorescent fusion proteins: random insertion of GFP with an in vitro transposition reaction." BMC neuroscience... This type of interference is known as steric hindrance. For instance, GFP may occlude catalytic sites...sites or motifs that are necessary for post-translational modifications. Despite these potential issues...targeted to the endoplasmid reticulum based on the appearance of OSER whorls. This assay is quantitative and...various fluorescent proteins (Botman et al, 2018; Cranfill et al, 2016; Lambert, 2019). These characteristics...applications, many were concerned with steric hindrance by the fluorescent protein. Thus, scientist began...

...variants (hereafter referred to as Cas9) and a single stranded guide RNA (gRNA) is sufficient to modify the genome...variants have been created in an effort to increase translation in a particular plant species or cell type (Belhaj...CRISPR components can be expressed stably or transiently depending on the delivery method and cell type...CRISPR components can be delivered and expressed transiently using a standard detergent, Polyethylene Glycol... More information on Agrobacterium-mediated transformation can be found in this blog post. The pDGE Dicot...Simplified schematic of agrobacterium-mediated transformation in plant cells. Agrobacterium tumefaciens can...plant genome. B) The Ti plasmid and helper are transformed into agrobacterium tumefaciens and exposed to...

...into smaller pieces, run on a gel, and transferred to a membrane for analysis through hybridization of ...researchers generating transgenic animals may choose Southern blotting to screen founder transgenic animals for ...techniques used to generate transgenic mouse lines, such as BAC transgenic or traditional knockins/knockouts... control sample. Southern blotting applications range from confirming the results of cloning or knockout...for the inserted transgene because Southern blots have a lower false positive rate than real-time PCR (4...hydroxyl group required for the extension of a DNA strand (8). Adding a small amount of a single radiolabelled...regular nucleotides results in the synthesis of DNA strands of all possible lengths because extension is terminated...

...proteins, which are directly trafficked to cell membranes. Membrane trafficking of farnesylated derivatives enables...Brainbow-3.0, but with mOrange2, EGFP and mKate2 as the fluorophores; mOrange2 is the default state. In... virus post-transcriptional regulatory element (WPRE), commonly used to increase transgene protein levels...fluorescent proteins: RFP (red), YFP (yellow) and M-CFP (membrane tethered cyan). Without Cre recombination, RFP...first fluorescent protein in the construct, OFP (orange), is expressed when Cre is absent; stochastic Cre...This system differs from Brainbow-1.0 in that transient Cre recombination is required to prevent constant... both fluorescence and sequence overlap (coral mOrange2, jellyfish EGFP, and sea anemone mKate2.) They...

...this FRET you speak of? Förster Resonance Energy Transfer (FRET) was originally described by Theodor Förster...alternative acronym Fluorescence Resonance Energy Transfer. Unlike the typical excitation and emission of...excited fluorophore, FRET involves a non-radiative transfer of energy (i.e. no emitted photons) from the excited...excitation by absorption of a photon 2. Energy transfer from the excited donor directly to the acceptor...the acceptor fluorophore. Otherwise the energy transferred from the donor will not be able to excite the...other and be oriented appropriately for energy transfer via dipole-dipole interaction. The efficiency ...biological processes occur within the typical FRET range, FRET efficiency is used to infer an interaction...

...concentration lies within the dynamic range of the curve. The dynamic range is the linear span between the lowest...the dynamic range of your standard curve, so if your sample does not fall within this range, it may be ... of the series dilutes by 1:2 for a series that ranges from 1:2–1:32. The specific dilution series used...Figure 1: A 1 mg/mL control is diluted in a series ranging from 1:2 to 1:32. Each individual step of the series... how well the data points fit the trendline. R2 ranges from 0 to 1 with 1 being a perfect fit and 0 indicating...that the curve can accurately measure.  Dynamic range depends on a number of factors, including the dilution...being used to measure the response. The dynamic range needs to be wide enough to cover all possible concentrations...

...members to the Board of Directors: Khandan Baradaran, Frank Stegmeier, and Luk Vandenberghe. We’re excited...members and get to know them! Khandan Baradaran, PhD Khandan Baradaran’s expertise in viral safety, gene therapy... with health authority regulations,” Baradaran says.  Baradaran’s path to science began in high school... getting these reagents is profound.” Frank Stegmeier, PhD Frank Stegmeier is the Chief Scientific Officer...in viral genetics. In regards to open science, Baradaran thinks the best way to solve problems is to work...think will get the research funded,” she says. Baradaran met Addgene Executive Director Joanne Kamens several...the targets that have the biggest potential to transform patients' lives. Previously Stegmeier was the ...

...process known as bioluminescent resonance energy transfer (BRET). The fluor then emits light at a tissue-penetrating... consists of NLuc fused to the more penetrative orange fluorescent protein fusion (OgNLuc). Both fluorescent...for excitation. The result was the cyan-excitable orange fluorescent protein, CyOFP1. CyOFP1 was molecularly...and improved maturation time compared to similar orange fluorescent proteins. In their work, the authors...great imaging tools generate cyan, yellow, green, orange and red fluorescence. The paper reports a wide ...Plasmid #85199), Yellow (YeNL; Plasmid #85201), Orange (OeNL; plasmid #85202) and Red (ReNL; plasmid #85203...have been made using fluorescent resonance energy transfer (FRET) probes. These probes generally consist ...

... affecting protein translation of different reporter proteins. Protein translation requires the start ...compared to CRISPR/Cas9 edits that rely on double strand DNA breaks. Due to this inefficiency it is crucial...constitutively expressed to identify successfully transfected cells. C-to-T editing by the APOBEC-Cas9 editosome...that results in the restoration of fluorescence. Transient reporter for editing enrichment (TREE) The Wang...Wang and Brafman lab developed a transient reporter for editing enrichment (TREE) to purify single base... to cells segregated solely by a reporter of transfection, which only report the efficiency of plasmid...frequency of base edits of interest than solely transfected cells. “Gene On” (GO)- a functional reporter ...

...72 hours post-transformation/transfection in a transient assay. For stable transformation, regenerated ...Agrobacterium-mediated transformation. We also constructed smaller vectors suitable for transient assays using ...changes a single base with an efficiency typically ranging from 25-75%, while the success of precise change...creates a nick in the DNA by cutting only a single strand, dramatically reducing the likelihood of detrimental...of the desired T-A base pair. Figure 1: Transition mutation (C>T, G>A, A>G, and T>C) installation...deaminases (ecTadA), fused it with dCas9, and transformed it into E. coli harboring the defective CamR ...using protoplast transformation. Cytosine base editors for plant There are several CBEs available for plant...

...attractive due to the ease of transduction in difficult-to-transfect cell types ranging from primary to embryonic... traditional transfection and random genomic integration of a plasmid. Viral transduction is also generally...integrated into the genome of the target cell or transiently expressed. It’s important to note that for all...vector can express your construct in target cells transiently or constitutively depending on the virus used...generally less toxic to cells than transfections, preserving cell viability. Library delivery Screening... entire genome, overexpression of all human transcription factors, barcoded libraries to trace cell lineage... with viral vectors instead of a traditional transfection method. This is because viral systems have a...

...Efficient gene transfer in C. elegans: extrachromosomal maintenance and integration of transforming sequences...easily manipulated with tools like CRISPR to make transgenic lines. The fruit fly (Drosophila melanogaster... in only 6 weeks compared to months to make a transgenic mouse line. Over the years the fly has become...DNA or RNA to edit their genomes or to create transgenic animals. Zebrafish are an ideal model organisms...development studies as their embryos are completely transparent. Thus scientists can easily observe early developmental...can be difficult in other vertebrate species. Transparency also allows scientists to easily observe fluorescently... control the temporal expression of different transgenes, the Stainer group determined that there are ...

...The development of the transient transfection platform for assaying transcriptional activity, for example...such as p300 and CBP histone acetyltransferases or CARM1 methyltransferase. This attendant signaling feature...dietary origin) that are diffused or actively transported from cells, traverse the body through the bloodstream...ligands NRs represent the largest family of transcription factors found in metazoans. The superfamily ... govern expression of genes involved in a broad range of reproductive, developmental, metabolic, and immune...multiple cell signaling systems. Activation of membrane receptors and signaling cascades conceivably allows...profiling datasets in to our data mining tool, Transcriptomine. We are currently nearing the completion of...

..., expressing Ad E1A and E1B, can be transfected with your transfer plasmid, carrying your gene of interest... performed 12–18 hours post-transfection to help remove toxic transfection reagents and keep the cells...helper plasmids, commonly called “triple plasmid transfection” (Figure 1).    Figure 1: Overview of key steps...cells happy. Harvesting virus Once you have transfected your cells, wait 2–5 days and then collect the supernatant...the concentration of viral particles that can transduce cells, which can vary based on target cells and... capsids, thus leading to an overestimate of transducible units in the virus preparation.  rAAVs  Now...Whitney, O., Walther, N., & Maurer, A. C. (2023). Transgene Expression in Cultured Cells Using Unpurified ...

...respondents (transfection method, amount of each component per transfection etc.). Take Pooran's NgAgo Survey...Doudna Lab Many thanks to our Guest Blogger Pooran Dewari! Pooran Dewari is a postdoc at the MRC Centre for...TOOL This post was contributed by guest blogger Pooran Dewari. Any views in this post are those of the...an ssDNA guide. Therefore, researchers must co-transfect cells with 5’-P-ssDNA guides and an NgAgo expression...delivery methods. Researchers can conveniently transfect/inject cells with in vitro preassembled Cas9/gRNA...). View the Complete and Up-to-Date Results of Pooran's NgAgo Survey The take-home message While most ...researchers will have to carry out repeated transfections with the ssDNA guide, although many may find...

...target and nontarget DNA strands upstream of the PAM site found in the nontarget strand. Protein Cas: ...sequences and keeping the array organized. Target strand: The strand of foreign DNA that Cas proteins recognize...recognize using an RNA guide. Non-target strand: The paired strand of foreign DNA cut by Cas9, whose sequence... and variable spacer sequences. The array is transcribed and processed into crRNAs, each including one...protospacer-adjacent motif” present in the non-target strand of DNA to be cut, which varies between CRISPR systems...from Addgene!   RNA crRNA: “CRISPR RNAs” are transcribed from the CRISPR array and include a spacer and...track down its suspect—the target DNA. tracrRNA: A “trans-activating CRISPR RNA” base pairs to the repeat ...

...between the edited strand and wild-type strand of the host DNA. When these strands hybridize, the nuclease...nuclease can detect strands with mismatches and cleave them. The results are then visualized using gel electrophoresis...is also key to perform these validations as translational researchers begin to use CRISPR as a therapy... search criteria are not exhaustive. Mismatch tolerance settings are often limited to off-target sites...nucleotides) that may still lead to off-target double-stranded breaks (Tsai et al. 2015). Additionally, current... genome-wide, unbiased identification of double-strand breaks (DSBs) evaluated by sequencing (GUIDE-seq...et al. 2013). GUIDE-seq and BLESS detect double-stranded breaks and do not require high sequencing read...

...different DNA strand comes along, it can attack this covalent bond thus joining the two DNA strands and releasing...planning to transform right away OR you can store the reaction at -20C overnight. 4. Transform TOPO Cloning...DNA sequence 5´-(C/T)CCTT-3' and digests double stranded DNA at this sequence. The energy from this breakage...stored as a covalent bond between the cleaved 3′ DNA strand and a tyrosyl residue of topoisomerase I (1). If...researcher to perform blue-white screening after transformation - self joining of the vector ends results in... minutes will not significantly improve the transformation efficiency).  5. Select and Analyze 10 White...add will depend on whether you are planning on transforming your reaction into chemically or electro-competent...

...scientists to quickly and easily generate zebrafish transgenic lines to study neurobiology and neuorodgenerative...pluripotent stem cells) through the expression of transcription factors OCT3/4, SOX2, KLF4, and c-MYC is the... method for generating iPSCs to express the transcription factors from a polycistronic cassette. However...comparable experimental results. Using PiggyBac transposons, the Woltjen Lab compared different variants ...reprogramming discrepancies. The polycistronic piggyBac transposon vectors tested in this work are available at ...approach Genetically modified organisms like transgenic mice are important tools for understanding the...a donor in FRET (fluorescence resonance energy transfer) and FLIM (Fluorescence Lifetime Imaging Microscopy...

...hiPSCs for clinical purposes: mRNA transfection- this allows transient expression of the reprogramming factors...cells, thus expressing additional EBNA-1 transiently. The transiently expressed EBNA-1 increases protein expression...insertional mutagenesis, which is the integration of transgenes into the genome that can cause disruption of ...express reprogramming factors To date, the mRNA transfection method has not been used successfully to derive...using PBMCs as the source of somatic cells. This translates to only about 16 hiPSC colonies per 100 ml of...such low efficiency of reprogramming is a major hindrance in using PBMCs as a source of somatic cells for...system thus provides a practical way to generate transgene-free, virus-free clinical grade hiPSCs from easily...