We narrowed to 846 results for: ins

...Carolina (plasmids at Addgene). It consists of two domains, LOV2 and Zdk1, where Zdk1 binds LOV2 with high...normally sits on microtubules to attract other proteins involved in their growth and extension. Now, they...domain. Each half was fused with one of the LOVTRAP domains. When both halves of the modified EB1 protein are...inactivated by the separation of its synergistic domains. This paper demonstrated that the migration of ...fluorophore binding aptamers and MS2-binding fluorescent proteins were the state of the art in RNA imaging in vivo...on the lookout for better, brighter fluorescent proteins (FP) with improved tissue penetrance and live ...

...Argonaute proteins are also present in prokaryotes, and some of these proteins protect against invading...reasons why you might choose NgAgo over CRISPR proteins Cas9 or Cpf1 - keep reading to learn more! Background...: Argonaute and NgAgo In eukaryotes, Argonaute proteins play an important role in RNA interference, binding...when the protein is being synthesized, so NgAgo remains faithful to its guide DNA and cannot switch guides...

...usage has been limited by technical problems. For instance, it’s much more difficult to introduce a gene ...strategy for transducing primary cells. What if, instead of supplying a DNA construct, they could simply... adapted from D’Astolfo et al. In certain instances, the potential benefits of nonviral transduction...with virus work. Fourth, working directly with proteins removes the limitations of packaging size associated...References Efficient Intracellular Delivery of Native Proteins. D'Astolfo DS, Pagliero RJ, Pras A, Karthaus WR...PubMed. Find the Plasmids @Addgene: iTOP-tested proteins and reporters Previous Protein Delivery Techniques...

...Tom, and Alan E. Tomkinson. "Eukaryotic DNA ligases: structural and functional insights." Annu. Rev. Biochem...many molecular biology protocols, including vector-insert joining for recombinant plasmid construction (restriction...DNA ligase, are naturally able to discriminate against ligating substrates containing base pair mismatches... can be achieved through a combination of the intrinsic selectivity of the ligase active site and careful...indicated that the optimal buffer for Taq DNA Ligase contains 100 – 200 mM KCl at pH 8.5. This optimization....6 (2004): 757-764. PubMed PMID: 15582400. 2. Tomkinson, Alan E., et al. "DNA ligases: structure, reaction...

...but can also be used to brainstorm on data in smaller groups, where for instance the presence of the PI...lab meetings with a focus on discussion and brainstorming Though, we might be forced to work from home...of each talk - time for free interaction and brainstorming. This could be achieved by asking each presenter...problem! You can still creatively collaborate and brainstorm! Image from Joseph Mucira.   Implement new... be written by several authors from different institutes, disciplines, and cultural backgrounds.” This...

...freeze-dried, cell-free (FD-CF) components. Making proteins outside of a cell - cell free extracts Typically...transformation), essentially giving the cell all the instructions it needs to produce that green fluorescent protein...systems contain all the parts needed to produce proteins without the need to handle a living, replicating...simply need to add water to express a variety of proteins. FD-CF reactions are also cost-effective, as each...easily sense, whether that’s bright fluorescent proteins that they can see, enzymes that can convert certain...activities fit your lesson plans. If you are an instructor at the college level, perhaps consider using ...

...accumulate inside the producer cells and purification protocols often describe a process that begins with cell... carried inside the vesicle, which may also contain other cellular molecules such as proteins or nucleic...cells also contain cellular macromolecules (e.g. proteins, nucleic acids) which possibly improve transduction... contexts, such as the delivery of therapeutic proteins to the brain which requires crossing the blood-brain-barrier...

...confirm the presence of an insert in a particular vector by excising the insert from the backbone. To do...use enzymes with restriction sites that flank the insert. You will need to know both the approximate size...vector backbone as well as the predicted size of the insert. You can search NCBI for YGOI to find the particular...right has a total size of 7.3kb, including a 1.2 kb insert. The plasmid was digested with 2 unique enzymes... and 1.2kb (red box), matching the backbone and insert, respectively. The results on the gel correspond...

...Associate Director of the Cancer Program at the Broad Institute of Harvard and MIT. The notion of cancer precision...We've developed a new plasmid toolkit, the Broad Institute Target Accelerator Plasmid Collection, that we...variant function and cancer phenotypes. Broad Institute Target Accelerator Plasmid Collections To overcome...this first challenge, we’ve developed the Broad Institute Target Accelerator Plasmid Collections, now widely...Associate Director of the Cancer Program at the Broad Institute of Harvard and MIT and is focused on making precision...Kits at Addgene Browse Plasmids from the Broad Institute ...

...Root from the Broad Institute and Alex Chavez from the Wyss Institute were instrumental in helping us establish...School, and Jonathan Ting’s lab from the Allen Institute have supported the AAV initiative from the beginning...Janelia Research Campus have provided valuable insight into AAV production. Finally, we are blessed to...permission from relevant plasmid depositors and their institutions to distribute viral preps derived from plasmids...

...restriction enzyme cloning uses short sticky ends to insert genes into plasmid backbones, base pairing between...absence of the other dNTPs. Once the vector and insert are (separately) digested to create 10–12 base ...on the power of homologous recombination. SLIC inserts and linearized vectors must have homologous DNA...imprecise T4 digestion (no dNTPs needed), allows both insert and vector to be digested in the same tube. Alternatively...these approaches require higher concentrations of insert to succeed. 5′ overhangs are generated in the linearized... be used with different types of PCR-generated inserts. By harnessing the power of DNA repair, you can...

...interact with the host. This is the role of Gag proteins: they contain the matrix, capsid, and nucleocapsid...assembly, and display. Pol, the last component, contains the reverse transcriptase and integrase components...convert the cargo from RNA to DNA and permanently insert the cargo into the host genome. Lentiviral-specific... Viral packaging and entry The viral packaging proteins for AAV are rep and cap. Rep is required for viral...discussed. LXs/Iva2 are primarily capsid assembly proteins required for packaging of the virus into particles...Resources Kulanayake, S., Tikoo, S. K. Adenovirus Core Proteins: Structure and Function. Viruses. 2021 13(3):388...

...proto-oncogenes and disrupt their expression (Robinson, 1982). To add insult to injury, retroviruses generally have...This viral DNA (also called proviral DNA) is then inserted into the host's genome and is used to make more...virus-specific elements such as the LTRs or structural proteins are not interchangeable between the two systems...Adenovirus generally has high immunogenicity, but some strains have been engineered to induce a lower inflammatory...immunogenicity low! References and Resources References Robinson HL. Retroviruses and cancer. Rev Infect Dis. 1982...

...if not methylated (if it comes from a phage for instance). Aside from restriction modification systems,...effectively cleave DNA. Common lab E. coli K12 strains such as DH5alpha contain 3 methylases that recognize...Dam or Dcm methylation, you can ensure this site remains unblocked by first cloning your DNA into a dam–...and re-purifying it. These specialized E. coli strains have been specifically engineered to be Dam and...those sites. Please keep in mind that dam-/dcm- strains may have an increased rate of mutation (as these...in mis-match repair functions of Dam), so these strains should not be used for long term storage. Additional...

...another name for a folder. For instance, your Desktop directory contains the folders and files that you...including background on other analyses that have inspired you to perform this analysis. Having a clearly...multiple projects in a given day. For example, instead of naming a file “output” name the file “1_output...subsequent word in a file name after the first word begins with a capital letter and words are not separated...temporarily used. Just be sure that the original code remains in tact! Take notes Comment in your scripts: ...

...small percentage of the proteins inside a cell have a commercial antibody against them. There’s also a question...Mendenhall of the University of Alabama in Huntsville explains it, a major goal in his laboratory is to understand...track down every last one of those gene regulatory proteins and the places in the genome where they can be.... If things work correctly, the second plasmid inserts a small epitope tag called FLAG at the end of the...

...and control genes, but according to the Broad Institute's Silvana Konermann, the first generation of CRISPR-Cas9...recognized as an ideal spot to fuse the activators instead of at the protein’s C terminus. With that new design... and knew she was on the right track. The next insight was to assemble a synthetic transcription activation...complex consisting of multiple distinct effector domains that work together synergistically, modeled after...CRISPRa) sgRNA library uses the sunCas9 system and contains 10 sgRNAs for each transcription start site in...The transcriptional repressor (CRISPRi) library contains 10 sgRNAs for each transcription start site in...

...occurring resistant mutants and the insensitivity of yeast strains to some antibiotics. Auxotrophy is ...bacteria, yeast can post-translationally modify proteins yet they still share many of the same technical...replication (Please note: This first section primarily pertains to ORIs in budding yeast, Saccharomyces cerevisiae...other hand, do not require a well defined ORI. Instead, the size and A-T content of the DNA (apparently...compound required for its growth. Many auxotrophic strains of yeast exist which can be easily maintained when...

...multiplexing of sgRNAs against unique genomic targets or multiple sgRNAs against the same target to ensure...protein purification and metabolic engineering. If instead you want to build genetic circuits and would benefit...library of parts but don’t need to add tags to your proteins, then the CIDAR Kit is an excellent option (Iverson...system includes a toolkit to express recombinant proteins within the mitochondrial matrix.     Figure...can be repurposed for CRISPR applications (gene insert = Cas9 + gRNA), Addgene distributes specialized...of a Toolkit for the Expression of Heterologous Proteins in Yeast Mitochondria.” BMC Biotechnology, vol...

...The Rare Genomics Institute BeHEARD (Helping Empower and Accelerate Research Discoveries) Challenge provides...our fourth year working with the Rare Genomics Institute by providing plasmid-based tools for rare disease...Addgene to set up assays to see what molecules or proteins are involved in IL-36 release in keratinocytes... valve (an aortic valve that has two leaflets, instead of three) and acute aortic dissections (a tear ...standing interest in the biology of GPCRs and G proteins, with an emphasis on oligomerization, signalling...Resources Find out more about the Rare Genomics Institute and this year's BeHeard Award! ...