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We narrowed to 920 results for: Gene

Showing: 1 - 40 of 920 results

  1. Plasmids 101: Gene, enhancer, and promoter trapping

    Type
    Blog Post
    Published
    May 2, 2023, 1 p.m.
    ...exon of a gene, and thus they do not disrupt the gene expression of the gene they monitor. Gene traps Gene...expression of a nearby gene. The reporter gene serves as a proxy for the endogenous gene – its expression levels...predicted genes have unidentified roles as well (Vanni, et al). Is it true that this many genes are truly...biologically relevant situations. Tissue-specific genes – the genes of interest may not be expressed in the tissue...are working with. Development-specific genes – if a gene functions at multiple stages in development... be observed. Functionally redundant genes – multiple genes which function in overlapping roles can...reporter gene flanked by a minimal promoter which can integrate proximal to an endogenous gene to turn ...

  2. Stabilized Bacterial Promoters: Constant Gene Expression at any Copy Number

    Type
    Blog Post
    Published
    Sept. 4, 2018, 12:53 p.m.
    ...of where the genes were expressed. What’s the use of stable gene expression? Constant gene expression can...Researchers express genes of interest from plasmids in order to study gene function or to engineer cells...input and gene expression is the output. The input, plasmid copy number, directly influences gene expression... varied in size or carried different genes. In these cases, gene expression also remained stable regardless...media and growth stage can both affect gene copy number. Moreover, gene copy number is affected differently...the levels of gene expression from stabilized promoters were equivalent whether the genes were expressed...level of gene expression. For example, the synthesis of deoxychromoviridans requires three genes whose proteins...

  3. Plasmids 101: Positive and Negative Selection for Plasmid Cloning

    Type
    Blog Post
    Published
    Aug. 22, 2019, 12:43 p.m.
    ...occurring genes for cloning purposes (antibiotic resistance genes, toxin genes, and the sacB gene). By tapping...gained a specific gene survive In positive selection, only cells that contain a specific gene survive. The ...selecting for the loss of a gene product - usually something toxic. This gene is found on the original plasmid...insertion of a DNA fragment within the gene or loss of the gene alleviates its toxic effect. Negative ...destination plasmid, the ccdB gene is flanked by recombination sites. To introduce a gene of interest into the ...be added to flank the gene. Successful recombination and integration of the gene of interest at these ...plasmid excises the ccdB gene and the cells will only grow if they contain the gene of interest. SacB counter-selection...

  4. Plasmids 101: Knockout/Knock-In Plasmids

    Type
    Blog Post
    Published
    Dec. 1, 2016, 3:30 p.m.
    ...targeted gene will be removed from the chromosome and and replaced by the resistance gene.The gene is thus...strategies to investigate a gene's function is to inactivate, or "knockout", the gene by replacing it or disrupting...endogenous gene. Here, we'll describe the techniques and the plasmids used to inactivate specific genes in mammalian...given gene.   Figure 1: A knockout targeting vector designed to insert a resistance gene. The ...targeted gene have been cloned into the vector on either side of an antibiotic resistance gene. To avoid... resistance gene through the insertion of flanking LoxP sequences, you can remove the gene through the...later remove a resistance gene. In this construct, the neomycin resistance gene is floxed so that it can...

  5. AAVs in Retinal Gene Therapy

    Type
    Blog Post
    Published
    July 31, 2018, 12:55 p.m.
    ...human genetics led to the identification of several disease-causing genes, including the RPE65 gene responsible...I got into the field of gene therapy, more specifically AAV-based retinal gene therapy. The year was 2001...clinical successes for gene (and cell) therapies. One may wonder why the first AAV-based gene therapy treatment...blindness) is caused by defects in the gene MYO7a, at ~7 kb this gene is too large to deliver via AAV.  Dual...Moving forward with ocular gene therapy The pipeline of AAV-based ocular gene therapies is impressive: ...and further reading 1. FDA approves gene therapy for blindness 2. Gene Therapy emerges from disgrace to ...the treatment of some genetic diseases. I didn’t know it at the time, but gene therapy had fallen - ...

  6. Adeno-associated Viruses (AAVs) for Genome Editing

    Type
    Blog Post
    Published
    March 27, 2018, 1:32 p.m.
    ...fantastic gene delivery vehicles for episomal gene expression and are particularly useful for gene delivery...downstream of a functional promoter (gene traps). The resultant gene targeting system could be used to routinely...tumor suppressor genes in human cells (Kim et al 2008). They have also used AAV gene editing to introduce...your gene of choice. These vectors are very similar, both with polylinkers for the addition of gene-specific...acceptor-IRES-neoR gene, which enhances the efficiency of homologous integration via gene trap enrichment...promoter-driven TK-neoR gene. To modify these tagging vectors for your specific gene, the lab recommends ...experiments, the floxed neoR gene is removed via infection with adeno-cre. AAV-based gene editing compared to ...

  7. Transgenic Organisms, Cas9 Gene Drives, and Appropriate Safeguards

    Type
    Blog Post
    Published
    May 22, 2015, 4:58 p.m.
    ...creating a gene drive? The simplest way to make a gene drive is to insert an endonuclease gene within its...endonuclease gene. As we pointed out last year2, delivering a DNA cassette encoding the cas9 gene and single...designed to overwrite a previous gene drive and thereby undo the genetic changes driven by the earlier intervention2...molecular biology and genetics because the underlying advance enabling RNA-guided gene drives is Cas9 genome...formal stance taken by Addgene or its staff. Additional information about gene drives can be found in ...Burt, A. Site-specific selfish genes as tools for the control and genetic engineering of natural populations...should be aware of the potential hazards of creating “gene drives” capable of spreading through wild populations...

  8. Natural or synthetic? How Addgene’s dataset reveals trends in biological innovation

    Type
    Blog Post
    Published
    Nov. 8, 2018, 3:47 p.m.
    ... natural gene and a synthetic version of that gene was a clear predictor of whether the gene is synthetic... of the genes deposited at Addgene were synthetic. But nine years later, over 20% of the genes deposited...whether a gene is natural or synthetic just by looking at its sequence. MIT collaboration for gene classification...predicts whether a gene is synthetic or natural with 97.7% accuracy. Classifying genes as natural or synthetic.... If a gene was moved exactly as is from one organism to a distantly related organism, the gene may not...: rare. “Nature has many genes where there are not rare codons and many genes where there are instances...Pfingstag, and Neil C. Thompson. "Gene synthesis allows biologists to source genes from farther away in the tree...

  9. Adeno Associated Virus (AAV) for Cell and Gene Therapy

    Type
    Blog Post
    Published
    Nov. 7, 2017, 1:59 p.m.
    ...Thrasher. "Gene therapy progress and prospects: gene therapy for severe combined immunodeficiency." Gene therapy...a single gene, it can potentially be treated by correcting the mutation or replacing the gene. A notable... great gene delivery tools? There are a variety of key properties that distinguish AAVs as gene delivery...and effective. Challenges for AAV gene therapy Although AAV-based gene therapies are promising, scientists...mediates gene targeting and enhances endonuclease delivery for double-strand break repair." Gene therapy...of disease pathogenesis will enable robust therapeutic outcomes. In contrast to other gene delivery tools...by guest blogger Harshana S De Silva Feelixge. Gene therapy technologies hold great promise for improving...

  10. Plasmids for Endogenous Gene Tagging in Human Cells

    Type
    Blog Post
    Published
    April 6, 2017, 1:02 p.m.
    ... Figure 1: Gene editing is the addition of a useful coding sequence (i.e. GFP) into a gene while leaving...leaving other features of the gene intact. Here, we show an example gene targeted for tagging with GFP at...allele, or gene copy. The coding sequence for GFP is transcribed and translated along with the gene, producing...tags into your gene of interest. Researchers have developed a variety of endogenous gene tagging systems...plasmids for introducing tags into your gene of interest. Endogenous gene tagging with fluorescent proteins ...tag a wide variety of genes with GFP. These plasmids are available through Addgene, and the stem cell lines...everywhere To find out more about gene edited hiPS cell lines generated using the Allen Institute for Cell...

  11. Custom CRISPR Screens & the Green Listed Software

    Type
    Blog Post
    Published
    July 11, 2017, 2:30 p.m.
    ...presenter then showed which genes they had generated knockout mice for (all of the genes involved as I recall... each gene is not targeted by only one gRNA, instead many different gRNAs are used for each gene. At the...screens Differentially expressed genes  As the costs for global gene expression analyses like RNAseq are...test which of the differentially expressed genes found in a gene expression dataset are causal for a phenotype...differentially expressed genes that are known drug targets, or all differentially expressed genes that code for ...describes which genes were included, and how many gRNAs were identified for each gene. This file can be... if genes that you asked for were not found in the reference library. It’s not uncommon that genes have...

  12. Sonic Hedgehog? Sleeping Beauty? Learn About the Genes Behind Addgene’s Conference Room Names

    Type
    Blog Post
    Published
    April 2, 2019, 1:44 p.m.
    ... particular genes into mammalian genomes and easily disrupt gene expression to study gene function. Because...scheme? Gene names! This is not surprising for Addgene. But the names we chose were not just any gene names...Tinman, and Bagpipe. How does a gene get its name? Often times, a gene gets its name based on what happens...repress the expression of genes primarily involved in development. These genes are known to play a role..., two genes important for mesoderm differentiation Now, let's turn tinman and bagpipe, two genes important... defective tinman gene fail to develop a heart (Bodmer, 1993). Bodmer named the gene after the Wizard ...two homeo box genes that determine cell fates in the dorsal mesoderm of Drosophila." Genes & Development...

  13. Plasmids 101: Multicistronic Vectors

    Type
    Blog Post
    Published
    Sept. 9, 2014, 8:20 p.m.
    ...could replace one of the reporter genes above with your gene or genes of interest. Some of the plasmids...reporter gene). Addgene's collection offers a variety of plasmids to express two or more genes, some of... that are expressing your gene, especially if you are studying a novel gene, is not always a straightforward... your gene of interest, scientists have instead developed novel methods to co-express your gene along ...with a reporter, such as a fluorescence gene or a resistance gene. These reporters allow you to easily screen...more of the genes replaced.  Additionally, 2A peptides could be PCR-cloned between your genes of interest...Co-expression of multiple genes is valuable in many experimental settings. To achieve this, scientists...

  14. Save Time with Transient Plant Leaf Transformations

    Type
    Blog Post
    Published
    July 25, 2019, 12:03 p.m.
    ...overexpressing a gene of interest in the plant. A common method for silencing genes Virus induced gene silencing...by replacing the genes on the T-DNA region with our gene of interest or a reporter gene so that the transformation...transformation. Choose good reporter genes There are many great reporter genes available and which one you choose...transformation success. Some reporter gene examples are below. The GUS gene (β-glucuronidase) with histochemical...Hikmet Budak. "Virus-induced gene silencing, a post transcriptional gene silencing method." International...stable cell line or plant is regenerated from the transformed tissue. Gene function is commonly studied...via Addgene. For us, transient leaf transformations are a great help in screening candidate genes and ...

  15. Mouse Modeling, Part 1: Genetically Engineered Mice

    Type
    Blog Post
    Published
    July 11, 2019, 1:26 p.m.
    ...  Inducible gene expression Another type of genetically engineered mouse model allows gene expression ...an additional gene incorporated into their genome. While transgenic mice have the new gene construct incorporated...viral vector containing the gene construct can be used to insert the gene of interest into a zygote or...insert the gene at a specific site within the genome using homology arms that flank the gene of interest...knockout mice include a reporter gene, which replaces the targeted gene and provides a way to track successfully...present at both ends of a gene in the targeting vector, creating the floxed gene segment. Crossing of mice...target gene is initially inhibited by the presence of a stop cassette upstream of the target gene in the...

  16. CRISPR Activators: A Comparison Between dCas9-VP64, SAM, SunTag, VPR, and More!

    Type
    Blog Post
    Published
    Oct. 6, 2020, 1:15 p.m.
    ...for gene activation experiments. In cases of multiplex gene regulation (activating multiple genes at once...proteins had to be re-engineered for each gene, making wide-scale gene activation seem next to impossible. ...construct, it exhibits modest levels of gene activation, with some genes experiencing around 2-fold activation... When targeting single genes, SAM consistently shows the highest levels of gene activation compared to...recruited per targeted gene. Performance SunTag performs better than first generation activators while showing...Prior to the discovery of CRISPR/Cas systems, gene activation across multiple loci was an arduous process...systems, however, greatly improved the simplicity of gene activation: rather than requiring protein engineering...

  17. New Optimized Genome-wide CRISPRko, CRISPRi, and CRISPRa Libraries

    Type
    Blog Post
    Published
    Oct. 4, 2018, 12:44 p.m.
    ...function of lowly expressed genes or genes that are more drastically affected by gene activation rather than...investigate gene function. CRISPR libraries can be used to knock out, inhibit or activate target genes by combining...CRISPRa) to regulate gene expression. Some CRISPR libraries contain many sgRNAs per gene to increase confidence...of a gold standard set of genes (1,580 essential and 927 non-essential genes). The Root and Doench lab...viability. In addition, high copy number genes may mimic viability genes as they are subject to an increase ...overexpression library identified several overlapping genes, most genes were unique to the individual libraries. These...a few sgRNAs per gene. These libraries provide more information while using less resources and are particularly...

  18. Quick Guide to Working with Drosophila Part 2: Controlling Gene Expression in Flies with Gal4/UAS

    Type
    Blog Post
    Published
    July 21, 2017, 12:48 p.m.
    ...transcription machinery to the site to induce gene expression. Thus, genes and siRNA encoded downstream of the ...lines and study gene expression in many different tissue types! Although putting a gene of interest directly...multiple genes with Gal4/UAS system One can also use Gal4 to drive expression of multiple genes under the... to knock down gene expression, rescue knock out mutations, and overexpress viral genes. Additionally,...in case you want to express one gene in one cell type and another gene in a different cell type (6–8)....yeast (1). It is a Drosophila geneticist’s main workhorse to turn genes on or off. Gal4 is a transcriptional...Perrimon, Targeted gene expression as a means of altering cell fates and generating dominant phenotypes...

  19. RNA Interference in Plant Biology: New Tools for an Old Favorite

    Type
    Blog Post
    Published
    Oct. 27, 2020, 1:15 p.m.
    ...Furthermore, gene knockout strategies cannot be used to investigate essential genes. Historically, gene silencing...while gene knockouts have long been considered the “gold-standard” for LOF studies, the high gene copy ...two non-exclusive gene silencing pathways: transcriptional and post-transcriptional gene silencing (PTGS...modifications of complementary gene sequences (Figure 1). As a result, gene silencing can be easily induced...phenotype discrepancies between gene silencing and knockout methods of the same gene. However, recent work has...that is required dsRNA complementary to your gene or genes of interest to trigger RNAi activation. A typical...complementary to your gene target as inverted repeats (Figure 1). With this technique, multiple genes can be simultaneously...

  20. Pooled CRISPR Libraries Offer Genome-Wide Control for Large-Scale Functional Screens

    Type
    Blog Post
    Published
    Feb. 24, 2015, 7:50 p.m.
    ...essentially any gene,” she says. The technical leap from mutating and deactivating a gene or genes to selectively...system also enabled them to turn on genes anywhere in the genome for gene control over a 1,000-fold range....control genes, but according to the Broad Institute's Silvana Konermann, the first generation of CRISPR-Cas9...CRISPR-Cas9 plasmids were not designed with gene activation in mind. “We had not managed to create a system...CRISPR-Cas9 activators that work well enough on any gene that they could be used by people with little bioengineering...Konermann said. “If you want to activate all 20,000 genes, it is not possible to test each one and make sure...you are really confident about working for most genes and individual targeting modules. Those two things...

  21. Viral Vectors 101: An Introduction to AAV

    Type
    Blog Post
    Published
    May 30, 2023, 1:15 p.m.
    ...interested in heftier genes, you can coinfect with multiple viruses to express genes or gene sets over 4.7 kb...capacity of ~ 4.7 kb, meaning that your gene of interest (or genes) need to be 4.7 kb or less to fit in ...expressing multiple small genes, you can design single rAAV vectors with multiple genes packaged in them and...cell-specific gene expression and viral infection. Cre or Flp dependent switches control gene expression ... transient gene expression in a particular cell type to CRISPR genome editing, optogenetics, and chemogenetics... AAV is delivery of genetic cargo to a host cell. In order to make room for a gene of interest, AAVs used...icosahedral capsid, that can be used to express genes of interest in cell and animal models. AAVs are ...

  22. New Tools Enable CRISPRa for Neuroscience Applications

    Type
    Blog Post
    Published
    July 2, 2019, 12:51 p.m.
    ...isoform-specific gene expression. Gene isoforms refer to mRNAs generated from the same gene but with different...isoform-specific gene regulation — at least, more straightforward than generating cDNA for each gene isoform individually...first ask yourself whether gene overexpression is necessary (instead of gene knockout or knockdown, for...tools for modulating gene expression in the central nervous system (CNS). To address this technological ...combinations of transcriptional activators to induce gene transcription at genomic sites targeted by specific...libraries. However, using CRISPRa to target individual genes can be relatively laborious because multiple plasmids...packaged into AAV for in vivo delivery. Multiplex gene targeting is also possible and is only constrained...

  23. Viral Vectors 101: Parts of the AAV Packaging Plasmid

    Type
    Blog Post
    Published
    Dec. 3, 2020, 2:15 p.m.
    ...two genes: the replication (rep) gene and the capsid (cap) gene. In wild-type AAV, these two genes are...with BioRender.com   Replication (rep) gene The rep gene encodes four proteins that are required for... is called cross-packaging.  Capsid (cap) gene The cap gene encodes three AAV viral capsid proteins VP1...which contains the AAV structural and packaging genes, the adenoviral helper plasmid which contains the..., freeing up space in the viral genome so that a gene of interest can be packaged into the virus instead...Next let’s take a closer look at the rep and cap genes. Figure 1. The RepCap plasmid and its products...and packaging: Rep78, Rep68, Rep52, and Rep40. Rep gene expression is often driven by the AAV p5 or p19 ...

  24. X-CHIME: Context Dependent Germline Knockout in Immune Cells

    Type
    Blog Post
    Published
    Jan. 16, 2024, 6:13 p.m.
    ...time-consuming, as it only changes one gene at a time. CRISPR-based systems enable gene knockout in immune cells in... to create mice with gene knockouts in their immune systems. However, many genes have different functions...combination with different genes. Having CRISPR systems for assessing genes in defined contexts would ... deposited at Addgene by Arlene Sharpe’s lab, allow for wide deployment of in vivo gene knockout in immune...Annotation of genes in immune cells typically involves the creation of germline knockout mice, which ...the constitutive, ubiquitous knockout of single genes in immune cells in vivo (LaFleur et al., 2019a and...Mouse Strain C-CHIME Knocks out a pair of genes pXPR_219 Violet-excited GFP (vex) Rosa26-Cas9...

  25. Sleeping Beauty Awakens for Genome Engineering

    Type
    Blog Post
    Published
    June 30, 2015, 2 p.m.
    ...both introduce genes into an organism’s genome (see figure) and to disrupt endogenous genes at the site ...first-generation transposase (as determined by transposition assays of antibiotic-resistance genes conducted...). In their Nature Genetics paper, the lab showed that efficiencies of stable gene transfer using SB100x... the rapid generation of stable transgenic cell lines which can robustly express your gene-of-interest... exciting tool for producing specific genetic mutations and gene disruptions without the use of viral ...transposase enables robust stable gene transfer in vertebrates. Nat Genet. 2009 Jun;41(6):753-61. Epub 2009... et al. A new approach to gene therapy using Sleeping Beauty to genetically modify clinical-grade T cells...

  26. Educational Planarian Tools for Teaching Developmental Biology Techniques

    Type
    Blog Post
    Published
    May 24, 2018, 1:20 p.m.
    ...planarian genes, we encourage you to deposit them back with Addgene and expand the number of genes that other...expression of specific gene expression. The 49 deposited plasmids enable students to study gene expression and...planarians. For example, the gene Piwi labels neoblast cells and the gene Prohormone convertase 2 (pc2...beta-catenin. Other gene targets do not require amputation to be visible. For example, odf2 gene knockdown results...and functions of key regeneration-related genes in four inexpensive and commercially available planarians...types and for viewing how gene expression changes through time or after genetic manipulation. The technique...Scientists routinely use techniques to alter gene expression or to label specific cells, but there are...

  27. Mobile-CRISPRi: Bringing CRISPRi to Diverse Bacteria

    Type
    Blog Post
    Published
    April 4, 2019, 12:53 p.m.
    ...antibiotic resistance genes. Genes that are targets of antibiotics are often essential genes which creates a.... ICEBs1 carries the genes needed for conjugation, but can also carry cargo genes, like Mobile-CRISPRi...if that gene is a target of an antibiotic. For example, let’s take a look at the essential gene folA which...targeted essential and non-essential genes with two guides per gene. The library was grown in glucose minimal...essential genes were less represented while bacteria containing guides targeting non-essential genes remained...does not disrupt gene function and is stable and active for greater than 50 generations. Figure ...tools scientists can use to study them. For example, gene knockdown with CRISPR interference (CRISPRi) has...

  28. Minigenomes - a Safe Way to Study Dangerous Viruses Like the Ebola Virus

    Type
    Blog Post
    Published
    March 17, 2016, 2:30 p.m.
    ...replication and viral gene transcription (6). The EBOV genome contains seven viral genes, each flanked by ...sequence (gene start) and a transcription termination/polyadenylation signal sequence (gene end). A minigenome...Figure 1). All viral genes are removed and replaced by a single (non-viral) reporter gene (e.g. firefly luciferase...luciferase or eGFP) flanked by the EBOV-specific gene start and gene end sequences. Thus, the minigenome contains...Cloning the viral genes in this plasmid would be more difficult compared to if each gene were encoded in...transcribe the reporter gene (e.g. firefly luciferase). The polymerase recognizes the gene start signal and ...of the reporter gene. The polymerase will elongate the mRNA until it reaches the gene end signal, where...

  29. Plasmids 101: The Promoter Region – Let's Go!

    Type
    Blog Post
    Published
    April 3, 2014, 8:05 p.m.
    ...depending on its orientation to your gene. T7lac High levels of gene expression Promoter from T7 bacteriophage...modulating gene expression through varied inducer concentrations. trp High levels of gene expression...human or mouse) General expression mRNA Mammalian promoter from phosphoglycerate kinase gene.  Constitutive...deacetylation. Ubc General expression mRNA Mammalian promoter from the human ubiquitin C gene Constitutive  ... human beta actin General expression mRNA Mammalian promoter from beta actin gene Constitutive  Ubiquitous... Requires the presence of Gal4 gene to activate promoter. Ac5 General expression mRNA Strong insect ...Actin 5c gene Constitutive  Commonly used in expression systems for Drosophila. Polyhedrin General expression...

  30. Quick Guide to All Things Lentivirus

    Type
    Blog Post
    Published
    March 21, 2017, 2:30 p.m.
    ...the env gene containing the rev response element (RRE), an internal promoter, and the desired gene (transfer...-1 genome containing all viral genes with the exception of the env gene (packaging plasmid), and 3) the...used to express your gene of interest either constitutively or inducibly. If your gene of interest is toxic...expression of your gene. The addition of Dox in the culture will then repress your gene of interest. This... Youichi Suzuki (2011). Gene Regulatable Lentiviral Vector System, Viral Gene Therapy, Dr. Ke Xu (Ed.)...harmful genes into a transfer vector. Lentiviral particles expressing proto-oncogenes or oncogenes for instance...safe use in a clinical setting. In this generation, the HIV Tat gene (previously used to drive expression...

  31. Advancing Biology with Zebrafish: Genetic Tools for Developmental Studies and More

    Type
    Blog Post
    Published
    Aug. 13, 2019, 12:58 p.m.
    ...70% of human genes have a counterpart in zebrafish. Understanding the function of these genes in fish will...unknown functions for genes, and lays the groundwork for understanding the gene networks that regulate...zebrafish-specific gene expression databases that provide information about when and where genes are active in...investigations. Genetic resources for zebrafish A common approach to unravel the gene networks that drive...model organism is its amenability to genetic manipulation. Tools for gene manipulation together with information...Additional mutagenesis projects have since been initiated and today thousands of mutations in genes affecting...other resources. Targeted gene deactivation tools for zebrafish Random mutagenesis screens provided a major...

  32. To Codon Optimize or Not: That is the Question

    Type
    Blog Post
    Published
    Nov. 12, 2020, 2:15 p.m.
    ...Codon optimization is an approach in gene engineering to improve gene expression by changing synonymous ... worth codon optimizing a human gene for expression in E. coli. Genes are also optimized for expression...ThermoFischers GeneArt Gene Synthesis NovoPro Labs Codon Optimization Tool (ExoOptimizer) Genewiz Codon Optimization...Slimko and Lester, 2003).  Addgene has a large selection of codon optimized genes, so check out our inventory... for a molecular biologist who wants to express genes from one organism in another? Let’s take a look ...correlated with codon bias across all endogenous genes in E. coli and S. cerevisiae (Tuller et. al., 2010...usage frequency in a reference of highly expressed genes (Sharp and Li, 1987). In 2000 the Ikemura lab created...

  33. Popular Retroviral Vectors and Their Uses in Scientific Research

    Type
    Blog Post
    Published
    July 24, 2018, 12:51 p.m.
    ...among other things) gene delivery, generating pluripotent stem cells, and developing gene therapies for monogenic...and lentiviral vectors that are used for gene regulation, gene of interest expression, knockout, and screening...Experimental uses include: Cre-dependent gene expression, gene knockout, and selection marker removal. ...Expressing your gene of interest pLenti-puro (Plasmid #39481) You can clone your gene of interest into...split-vector systems’ by separating viral genes and transgene expression across several plasmids. The most... host genome and thereby enable long-term stable gene expression. Gamma-retroviral vectors are derived...diseases. Additionally, retroviral vectors can regulate gene expression by delivering shRNA or miRNA. The ever...

  34. Rett Syndrome: A History of Research and Therapeutic Outlooks

    Type
    Blog Post
    Published
    Nov. 19, 2020, 2:15 p.m.
    ... on therapeutic strategies that use gene replacement therapy, gene editing, MECP2 reactivation, RNA editing...Novartis and rebranded to Novartis Gene Therapies, and Taysha Gene Therapies, founded on Gray’s work, ...protein 2 (MECP2) gene, Rett Syndrome is a rare, progressive, neurodevelopmental genetic disorder that affects...of Rett Syndrome research, the role of the MECP2 gene, and highlights some of the latest breakthrough ...painstaking work trying to isolate the causative gene for Rett Syndrome, Huda Zoghbi's lab discovered ...the first disease-causing mutations in the MECP2 gene leading to Rett Syndrome (Amir et al., 1999). We...global transcriptional regulator of thousands of genes. Studies have suggested roles in transcriptional...

  35. Illuminating Epigenetics with A FRET Based Biosensor

    Type
    Blog Post
    Published
    Nov. 19, 2015, 3:30 p.m.
    ...with the genes in the compacted region and results in a loss of gene expression. In this way, genes in compacted...the corresponding gene products due to the same region, but not the same exact genes, being improperly...? What exactly is epigenetics? Epigenetics is the study of heritable changes in gene expression that do...makes  genes less transcriptionally active. The role of epigenetics in studying disease Epigenetic changes...your genes documentary PBS Additional Resources at Addgene Find the Ting Lab plasmids at Addgene Browse... change in genotype. Epigenetic modifications play a large role in turning genes ‘off’ and ‘on’, signaling...fascinating examples of epigenetics in the form of imprinting. In imprinting, one copy of a gene (either the maternal...

  36. Genome-wide Screening Using CRISPR

    Type
    Blog Post
    Published
    Aug. 18, 2015, 2:30 p.m.
    ...trying to target every gene in the genome, or a specific class of genes? Addgene currently carries several...within the target gene. CRISPR can also be used to activate or repress target genes without permanently... gRNAs targeting the same gene can be strong evidence that a particular gene is actually important for...2) What genetic modification are you trying to make? Addgene carries CRISPR libraries for gene knockout...which the underlying genetic cause is not entirely known. Identifying which genes are important for a ... experiment is to generate and screen a population of mutant cells to identify genes involved in a particular...Currently, Addgene carries CRISPR libraries that target mouse, human, fly, E. coli, and T. gondii genes.  2...

  37. CRISPR Activation: A Practical Guide

    Type
    Blog Post
    Published
    Aug. 18, 2016, 2:30 p.m.
    ...prevent your gene from being up-regulated. For instance, generally, the more highly expressed a gene is under... research CRISPRa for specific gene targets CRISPR activation for genes with known functions has the potential...uses for Cas9 activators is in genetic screening. gRNAs targeting every gene in the human genome for example...multiple gene loci simultaneously, CRISPRa can also be used to map molecular pathways and genetic interactions...promoter of another gene, thereby driving aberrant transcription. That being said, we generally pick guides by...constructs allow for the upregulation of essentially any gene by fusing mutants of Cas9 deficient in DNA cutting...potential to be used in therapeutics, as it can increase gene expression at certain loci to create changes in ...

  38. Hot Plasmids - June 2019 - Optogenetics, Acoustic Reporter Genes, microRNAs, and the CRISPR-Cas9 system CHIME

    Type
    Blog Post
    Published
    June 6, 2019, 12:41 p.m.
    ...developed four acoustic reporter gene (ARG) plasmids to better monitor gene expression in vivo using ultrasound...plasmids at Addgene. Liu Q et al., Neuron. 2019. PubMed PMID: 30704911. Acoustic reporter genes for noninvasive...by Lukas Morgan  Listen to the acoustic reporter gene podcast segment In recent years, scientists are ...through a host organism, the expression of some genes can change depending on the location. Current methods...of microbes rely heavily on fluorescent protein genes. These optical reporters have limited deep-tissue...ultrasound. These ARG plasmids contain genes for “gas vesicles,” a gas-filled protein structure that water-bound...demonstrates engineered gas vesicles as the first reporter genes using ultrasound and provide a major benefit to ...

  39. Plasmids 101: Codon usage bias

    Type
    Blog Post
    Published
    Sept. 27, 2018, 1:09 p.m.
    ...highly expressed genes (Emilsson and Kurland, 1990). Controlling gene expression through gene sequence - Proteins...because you optimize a gene with one of these tools, it doesn’t necessarily mean the gene is going to express...5. Maertens, Barbara, et al. "Gene optimization mechanisms: A multi‐gene study reveals a high success ...getting your genes of interest codon optimized by ordering plasmids containing them from Addgene. If a plasmid...plasmid at Addgene contains a gene that’s been codon optimized for a particular organism, this will sometimes...available from Addgene now have full sequence data, we recommend directly analyzing gene sequences for ...changing conditions - Organisms often need to express genes at different levels under different conditions. ...

  40. pSiM24: Simplifying Plant Genetic Engineering

    Type
    Blog Post
    Published
    Sept. 29, 2015, 2:30 p.m.
    ...ideal for large gene transfer. The M24 promoter provides constitutive, high-level gene expression and ...further customizing gene expression. Sahoo et al. have tested the system with genes up to 4 kb in length...versatile gene expression vector for transient assays as well as stable expression of foreign genes in plants...Scientists have worked to optimize this system for gene transfer, studying the stability of modified Ti plasmids...versatile binary Ti vector for both transient and stable gene expression applications in plants. This smaller,...and many plant scientists want to transfer large genes, smaller binary vectors with a larger cloning capacity.... They also replaced the promoter regulating the gene of interest with a constitutive, robust M24 promoter...
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