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We narrowed to 741 results for: REV

Showing: 141 - 160 of 741 results

  1. The Crispr Babies Saga Shows the Need for Action, Not More Delays

    Type
    Blog Post
    ...was to be revisited at meetings when sufficient time, additional public fora for data review, and more... Bioethics. On November 25, the MIT Technology Review dropped a bombshell report. A scientist working... were born in a twin live birth several weeks previously. While this claim has yet to be independently...report on the ethics of “Novel techniques for the prevention of mitochondrial DNA disorders.” This document...public views on emerging IVF techniques designed to prevent the transmission of mitochondrial disease;” The...Council began their research into both novel and previously established ethical issues raised around mitochondrial...stop shuffling our feet on necessary actions to review and decide on the future of genome editing technology...

  2. How Addgene Creates Content

    Type
    Blog Post
    ...guide always gets an expert review, separate from the author, and a second review from someone less familiar...channel just for asking people if they can write or review things and another for asking specific science ...answers from our incredible scientists. Expert reviews on blog posts are done on a case-by-case basis,...case of the antibody guide, primary research and previously written Antibody 101 blog posts were used, as...panel recommendations. A few discussions in the review process led to quick dives back into the literature...sharing repositories. Then we format, do a final review to catch those pesky typos, and hit publish. Making...content is an iterative process. We are constantly reviewing the resources we currently offer to identify gaps...

  3. Optogenetics + CRISPR, Using Light to Control Genome Editing

    Type
    Blog Post
    ...off-target activity, and prevent cytotoxic effects. For a more detailed review, check out our blog post...externally control the location, timing, and reversibility of the genome editing process. Read on to learn...expression while imparting spatiotemporal control, reversibility, and repeatability. The system developed by ...ASCL1 gene could be spatially controlled in a reversible and repeatable manner. With their light-activated... after 30 hr. The system was also shown to be reversible and repeatable when blue light (450nm) was cycled...paCas9-2 system could be spatially controlled and reversibly activated by turning blue light on and off. As...repressor (dCas9). The activity of all variants was reversible and repeatable. Since these initial studies ...

  4. A Practical Guide to Optimizing AAV DIO and FLEx Vector Expression

    Type
    Blog Post
    ... sites in the same orientation preventing the transgene from reverting. Figure created with BioRender.... (FSF) sequence prior to an ATG start codon to prevent transcription in cells where the recombinase was...direction of arrows) the inverted transgene is reverted into the sense orientation and one of the unused... diagram showing the mechanism of spontaneous reversion during AAV plasmid amplification. On the left ...cre-recombinase sites lead to greater frequency of reversion and leaky expression. On the right non-homologous...scrambled recombination sights lead to reduced reversion and less off-target expression. Figure created...2020) it was determined that the optimal titer to prevent off-target expression of the sensitive FLEx-split...

  5. Controlling for Off-target Effects with a New Genome-wide CRISPR Screen Design

    Type
    Blog Post
    ... and identifying >88% of previously identified essential genes while previous Cas9 and shRNA library screens...and non-essential genes for both pathways had previously been identified (Hart et al, Bassik et al). The...10% false discovery rate and identified 67% of previously identified ricin toxicity genes. The screen also... also discovered several genes previously not associated with ricin regulation, including almost all genes...Results from the screen also reproduced several previously observed characteristics that influence gRNA ...A systematic mammalian genetic interaction map reveals pathways underlying ricin susceptibility." Cell...to conduct genome-wide screens with CRISPR/Cas9 Review the basics of CRISPR with this cheat sheet Read...

  6. New Optogenetic Tools for Cytoskeleton and Membrane Control

    Type
    Blog Post
    ...structures, and genetic mutations are not easily reversible. Late last year, the Akhmanova Lab published ...This design allows highly localized, rapid and reversible microtubule severing and has very low dark-state...Mahlandt et al., 2023). These Opto-RhoGEFs provide a reversible and non-invasive way to activate a RhoGTPase ...light-induced binding of the plasma membrane. In previous publications, they described tools to control ... fusion protein away from its usual location, preventing it from carrying out its proper function (Figure...with these CIB1-Rabs in other contexts are sure to reveal even more insights. Super-duper lipid editor And...composition of the membrane itself? The Baskin Lab previously published OptoPLD, an optogenetically controlled...

  7. Viral Vectors 101: Systemic Capsids

    Type
    Blog Post
    ...Cells. Annual Review of Neuroscience. 2022 Jul 8;45:447-469. https://doi.org/10.1146/annurev-neuro-111020... Cells. Annual Review of Neuroscience, 45(1), 447–469. https://doi.org/10.1146/annurev-neuro-111020-100834...The blood-brain-barrier (BBB) exists to prevent substances in the blood, like proteins and viruses, from...interest, while miRNA target sites can be used to prevent expression in cell types not of interest. Systemic...both mouse strains. It is therefore important to review the literature for use of a capsid in your strain...al. 2022. Image created with BioRender.com. Abbreviations: AAV, adeno-associated virus; GRE, gene regulatory.../ENEURO.0373-21.2022 Brown, D., Altermatt, M., Dobreva, T., Chen, S., Wang, A., Thomson, M., & Gradinaru...

  8. Plasmid Cloning by PCR

    Type
    Blog Post
    ...TAAGCAGAATTCATGTGGCATATCTCGAAGTAC-3'. For the Reverse Primer, the design is similar, but we need to use the reverse complement to get...primer and the downstream restriction site to the reverse primer. Next, we need to examine the DNA sequence...homology to the ORF). We now need to generate the reverse-complement of this sequence so that we can successfully...successfully amplify the ORF. You can generate the reverse-complement using existing software (a quick internet...others). If we put the sequence we chose for our reverse primer (5’-TGGCATATCTCGAAGTACTGAGCGGCCGCTAAGCA-...TGGCATATCTCGAAGTACTGAGCGGCCGCTAAGCA-3’) into this calculator we get a final Reverse Primer sequence of 5’-TGCTTAGCGGCCGCTCAGTACTTCGAGATATGCCA...

  9. Plasmids 101: Screens vs. Selections

    Type
    Blog Post
    ...you. Screen variations Forward and reverse screens Forward and reverse screens involve manipulating or ...manipulation of the population. This is contrary to reverse screens, where there is removal of known factors...genes. This is a perfect example of an unbiased, reverse screen – all known genes are assayed for their ...types of screens are very similar to forward and reverse screens, except that they begin with a uniform ...manipulations which either enhance or suppress the previously defined disease phenotype. These screens employ... same downstream manipulations as a forward or reverse screen (CRISPR, drug, etc.), just with an added...

  10. The Many Reasons for Irreproducible Research- and a Vaccine to Eradicate It

    Type
    Blog Post
    ...is difficult to replicate.  Why is replicating previous research findings so difficult to do?  There are... so it is unlikely to be caught during the peer review process, which often focuses more attention on ...between the original setting and the new one, previously unaccounted for, may lead to different results...finding we can be more certain that some important, previously unknown variable actually makes a big difference...of Open Science. With a Registered Report, peer review occurs before results are known. The author submits...journals before a study is conducted. Editors and reviewers recommend changes, and if everyone agrees that...proposed experiment. Furthermore, the journal and reviewers are incentivized to make sure that any possible...

  11. 3 Challenges in Plant Synthetic Biology

    Type
    Blog Post
    ... Learn more about Revolution Bioengineering Follow RevBio on Twitter Like RevBio on Facebook Read...Keira Havens, co-founders of Revolution Bioengineering. Read their previous blog post about how they started...the public in an open, responsible manner. At Revolution Bio, we are addressing this challenge directly...project. We’ve shared some of the concerns that Revolution Bio had to address as we developed the color-...

  12. 9 tips for a successful postdoctoral experience

    Type
    Blog Post
    ... tips for a successful postdoc fellowship. 1. Reverse engineer success On Day 1 of your postdoctoral ...these goals. Planning is often more easily done in reverse. Assuming you want to become competitive for a ... typically takes 6-12 months depending on the revisions and the amount of time spent shopping a manuscript...conversations with senior investigators. Discussions can reveal mutual scientific interests, complementary perspectives...more positive grant application and manuscript reviews, etc. You'll also meet future collaborators and...impressions on potential manuscript or fellowship reviewers, expose your research to search committees, and...Seek opportunities to write manuscript drafts, reviews, cover letters, fellowships, etc. Second, get constructive...

  13. Uncertainty about Labor Law Brings More Uncertainty to Postdoc Wages

    Type
    Blog Post
    ...decisions to alter postdoc salaries - in some cases reversing these decisions entirely. At Future of Research...resource leading up to this date, as discussed in a previous Addgene blogpost and more recently in a paper ...initially cancelled their plans, but have since reversed their position and decided to allow individual... in Boston also just announced that they were reversing their plans to cancel raises). A further 9 institutions...What will these further changes mean? The abrupt reversal in salary raises at institutions will likely have...where salaries had already been raised higher previously and all 6,000+ postdocs at the University of ...Additional Resources on the Addgene Blog Read Gary's Previous Article: Changing Labor Laws Bring Increased Postdoc...

  14. Viral Vectors 101: The Retroviral Lifecycle

    Type
    Blog Post
    ... have its structural proteins expressed from a reverse-transcribed piece of DNA called a “provirus.” This...target cell” (or host cell) by the viral protein reverse transcriptase (RT). In the case of a natural virus...virus, both the protein and the genetic code for reverse transcriptase are packaged (Wu et al., 1997) inside...vector! For viral vectors, only the protein for reverse transcriptase (RT) is present in the capsid, as...translation of proteins because they integrate this reverse-transcribed DNA into the host cell’s genome and... in step 2 during uncoating. After uncoating, reverse transcription follows the release of the two strands...encoded by complex retroviruses. Microbiological Reviews, 56(3), 375–394. https://doi.org/10.1128/MR.56.3.375...

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

    Type
    Blog Post
    ...reporter gene transcription by the EBOV polymerase (reviewed in 2, 5). The minigenome is capable of being replicated... many nonsegmented negative sense RNA viruses (reviewed in 2, 3). Other important components of the Zaire...replication and transcription: NP, VP35, VP30, and L (reviewed in 5). The RNA genome is encapsidated by the nucleoprotein...arguing against this approach. First, this would prevent the study of the proteins independently from each...Central PMCID: PMC103829. 2. Conzelmann, K. K. "Reverse genetics of mononegavirales." Biology of negative...negative strand RNA viruses: The power of reverse genetics. Springer Berlin Heidelberg, 2004. 1-41. PubMed...replication competent virus-like particles and beyond: reverse genetics systems for filoviruses and other negative...

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

    Type
    Blog Post
    ...individuals with Rett syndrome experience regression of previously acquired skills such as speech, coordination,...landmark paper in Science showed that dramatic reversal of disease symptoms in mice is possible (Guy et... halted the neurological damage but completely reversed it, regardless of age or symptom severity. This... exciting finding injected hope that a similar reversal may be possible for children and adults with Rett...RNA trans-splicing, and protein replacement to reverse the impacts of this disease.  As a non-degenerative...non-degenerative, monogenic disease that has shown reversibility of symptoms in animal models, Rett syndrome is ...Guy J, Gan J, Selfridge J, Cobb S, Bird A (2007) Reversal of Neurological Defects in a Mouse Model of Rett...

  17. Antibiotic Resistance: An Old Solution but a New Problem

    Type
    Blog Post
    ... any preventive measure designed to help keep you from getting sick can be helpful in preventing the spread... on preventing the spread of antibiotic resistant bacteria, check out this resource on preventing antibiotic...because ‘antibiotic’ merely means that the drug prevents the growth of a living organism. As for multi-...standpoint, we can all work together to try and prevent outbreaks of drug resistant bacteria within communities...standards. Handwashing is an essential component of preventing infection, as is cleaning and disinfecting frequently...

  18. 28 Hot Plasmid Technologies from 2015

    Type
    Blog Post
    ... and the promoter to prevent leaky expression of this resistance. Unlike previous systems, pUCXKT does...can do so in a very specific, precise manner. Previously, scientists have been successful in regulating...enzymes (e.g. nucleases, recombinases) which irreversibly excise or invert a specific chunk of DNA. However...deposited a set of 11 phage integrases which act irreversibly on their cognate attB/attP sites with no cross-talk...subcellular localization in mammalian cell culture and reversible control of signalling pathways. These LIDs are... powerful tools which allow you to easily and reversibly switch off and on your favourite signalling pathways...the MCS has several options) and a pUCX-specific reverse primer containing the missing codons from the antibiotic...

  19. FLEx Technology and Optogenetics: Flipping the switch on gene expression with high spatial and temporal resolution

    Type
    Blog Post
    ... have taken researchers to the threshold of a revolution in modern neuroscience, an era in which technologies...antisense orientation with respect to the promoter to prevent its expression, and is flanked (“floxed”) by two...electroencephalography evidences of arousal in mice previously subjected to steady-state general anesthesia ...behaviors. The high spatio-temporal precision and reversible modulation of FLEx vectors combined with the ...functions, and molecular mechanisms." Chemical reviews 114.1 (2013): 126-163. PubMed PMID: 24364740. PubMed...development and application of optogenetics." Annual review of neuroscience 34 (2011). PubMed PMID: 21692661...of cre-loxp site-specific recombination." Annual review of biophysics and biomolecular structure 30.1 (...

  20. Hot Plasmids - September 2019 - CRISPR, Auxin Inducible Degradation, MerMAID Channelrhodopsins, and Genome Looping

    Type
    Blog Post
    ...plant hormones called auxins to conditionally and reversibly degrade proteins of interest fused to a degron...2009, Natsume et al., 2016).  The Kanemaki lab previously developed a CRISPR/Cas9 based method to tag endogenous... be used to engineer specific sites that were previously difficult. Find SpCas9 and Target-AID-NG plasmids...promoter and an enhancer. What makes LADL unique from previous iterations is the addition of CRY2, another Arabidoposis...hours of blue light induction (6-fold faster than previously developed systems). The LADL system promises ...it’s briefer time scale and its inducible and reversible nature. Find the LADL plasmids at Addgene Kim...
Showing: 141 - 160 of 741 results