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We narrowed to 605 results for: RAN-1

Showing: 101 - 110 of 605 results
  1. PiggyBac-ing Through the Genome Editing Field

    Type
    Blog Post
    Published
    May 31, 2016, 3:30 p.m.
    ...Correct Path.,” J. stem cell Regen. Biol., vol. 1, no. 1, Jan. PubMed PMID: 26702451. PubMed Central PMCID... 2005 [1]. Like other transposons, piggyBac™ has two components, a transposon and a transposase. The piggyBac...integration [8].   Figure 1: Mechanism of the piggyBac™  transposase/transposon gene modification system...limit [1]. The piggyBac™ technology can be used for numerous applications, including transgenesis, gene-trap...The predicted frequency of ‘TTAA’ in the genome is 1 in every 256 base-pairs of DNA sequence, making it...technologies and strategies for their projects. References 1. L. E. Woodard and M. H. Wilson, “piggyBac-ing models...titrating the transposase to transposon ratios. High ratios of transposase to transposon result in a greater...
  2. Viral Vectors 101: Virus Safety

    Type
    Blog Post
    Published
    June 22, 2023, 1:15 p.m.
    ...including yourself as the researcher.   Figure 1: Separation of retroviral packaging components onto...these reasons, AAV is usually classified as a BSL-1 safety level. Adenovirus is similar to AAV in that...lab coats are either recommended or required in BSL-1 and required in BSL-2 labs. When working with lentivirus...required whenever there’s a splash or spray risk in BSL-1. In BSL-2 labs, eye protection is required any time...isopropyl alcohol, 0.5% peracetic acid, 10% bleach, or 1% iodine are useful. Some organizations or cities may...looks like the genetic cargo in one plasmid (the transfer plasmid), the envelope (Env) gene on a separate... plasmid). Viral production then proceeds by transfection of all three plasmids into a producer cell line...
  3. Plasmids 101: FLEx Vectors

    Type
    Blog Post
    Published
    April 28, 2016, 2:30 p.m.
    ...coding sequence in the antisense orientation (Figure 1, top). The entire DNA cassette would be flanked by...include both Cre and FLP recombinases.   Figure 1 (modified from Schnutgen et al. Nat Biotech 2003):...identical sites on one end of the DNA fragment (Figure 1, middle). A second recombination event then excises...between the identical loxP or lox511 sites (Figure 1, bottom). Since only one loxP and lox511 site will...genOway press release for more information. References: 1. Schnütgen F, Doerflinger N, Calléja C, Wendling O...sites directs the Cre recombinase to invert, translocate, or excise a DNA fragment. The availability of...infected and would have the potential to express the transduced reporter gene. Generating a FLEx switch to control...
  4. Some Like it Hot: Thermostable GeoCas9

    Type
    Blog Post
    Published
    Sept. 14, 2017, 12:40 p.m.
    ...buffer at temperatures ranging from 25-80 °C. Activity is expressed as Kcleave (min-1). SpCas9 has maximum...Cells? Yes Yes Longevity in Serum?* No Yes Table 1. Comparing SpCas9 and GeoCas9. *For in vitro Cas9 ...GeoCas9 is active at high temperatures Figure 1: Thermostability of SpCas9 vs GeoCas9. SpCas9 or GeoCas9...outperformed SpCas9 at high temperatures. See Figure 1 for a summary of the experimental results. GeoCas9...). G. stearothermophilus grows at temperatures ranging from 30-74 °C, with an optimal growth temperature...or G) Spacer Length 20 nt 22 nt Temperature Range for Optimal Activity 35 - 45 °C 50 - 70 °C Genome...Harrington et al tested GeoCas9’s editing skills by transfecting HEK293 cells with GeoCas9 or SpCas9 RNP’s and...
  5. Improving Transduction Efficiency with Exosome AAVs

    Type
    Blog Post
    Published
    March 14, 2019, 1:54 p.m.
    ...vector-exosomes), exosome AAVs, or exo-AAVs (Figure 1). The researchers discovered that these particles ... are often found with cellular exosomes, small membrane vesicles or microvesicles (approx. 40 - 150 nm... diameter) that are secreted from the cellular membrane into the culture medium. AAV particles are either... vectors. They found that they have a higher transduction efficiency, i.e. the same number of AAV particles...to modify it to improve yields. For example, transfecting the producer cell line with an expression vector...proteins, nucleic acids) which possibly improve transduction properties. György and colleagues found that...that the association of AAV particles with membrane vesicles protects them from antibody recognition and...
  6. The Importance of High Titer for AAV Transductions

    Type
    Blog Post
    Published
    March 1, 2018, 2:11 p.m.
    ...methods to improve AAV production.     References 1.Clark, K. Reed. "Recent advances in recombinant adeno-associated... The higher the titer, the better. AAV transport and transgene expression It’s clear that we need high...be required. For example, retrograde AAV gets transported up the cell’s axon. While the mechanism for this...yet fully understood, we can imagine that this transport is probably not 100% efficient. So the few AAV... higher retrograde AAV titers for successful transduction. In another example, neuronal gene expression..., fewer channel proteins are required for the transduced cell to become light responsive than fluorescent...process of generating viral particles. Limit tube transfer - the fewer opportunities your viral particles...
  7. CRISPR 101: Non-Homologous End Joining

    Type
    Blog Post
    Published
    April 16, 2015, 3:45 p.m.
    ...842 . https://doi.org/10.1016/j.molcel.2014.08.002 Ran FA, Hsu PD, Lin C-Y, Gootenberg JS, Konermann S, ...Last updated Sept 1, 2020. This post was contributed by David Wyatt and Dale Ramsden, UNC at Chapel Hill...the course of repair by NHEJ are typically small (1-10 bp) but extremely heterogeneous. There is consequently... single strand break; when two such breaks are introduced near each other, in opposite strands. This “...Cas9 can be easily programmed to make a DNA double strand break (DSB) in the genome wherever the user chooses....org/10.1371/journal.pgen.1000683 Bétermier M, Bertrand P, Lopez BS (2014) Is Non-Homologous End-Joining...Giovannangeli C, Jasin M, Brunet E (2014) Chromosomal Translocations in Human Cells Are Generated by Canonical Nonhomologous...
  8. Viral Vectors 101: Pseudotyping

    Type
    Blog Post
    Published
    April 20, 2021, 2:30 p.m.
    ...tail.   Figure 1: Lentiviral production uses three plasmids: (1) The transfer plasmid, (2) the envelope... 2017)  Table 1 (Gutierrez-Guerrero et al., 2020) Table 1 (Cronin et al., 2005)     References and ... on different viral membranes, tropism, and advantages and disadvantages: Table 1 (Joglekar et al., 2017... specific purposes as described previously (Table 1, Gutierrez-Guerrero, et al, 2020). Because retrovirus...lentiviral vector to be transported retrograde and also has higher neuronal transduction efficiency and a better...virus to either restrict or broaden the host cell range (tropism). Pseudotyping is only done on enveloped...plasmid, and (3) the packaging plasmid. These are transfected into target cells which produce lentiviral particles...
  9. Tips and Tricks for Using Golden Gate Modular Cloning (MoClo)

    Type
    Blog Post
    Published
    July 8, 2025, 1:15 p.m.
    ... process. Also refer to Figure 1 for a visual overview! Figure 1: General overview of the MoClo... multiple colonies after each transformation. This could be as few as 1–2 colonies if you’re an experienced...part from an Addgene MoClo kit, to construct a Level 1 vector with a promoter and terminator flanking the...acceptor (bottom) for CDS1 parts. (B) Example Level 1 assembly. The choice between different Level 0 vectors...properly. Do this sequentially for your Level 0, Level 1, and Level 2 assemblies using the plasmid sequence... after each transformation and/or use different E. coli strains with higher transformation efficiency....vector with two other independently regulated transcription units. Created with BioRender.com.   Planning...
  10. Controlling for Off-target Effects with a New Genome-wide CRISPR Screen Design

    Type
    Blog Post
    Published
    Sept. 13, 2018, 1:55 p.m.
    ...holds true in genome-wide screens.   Figure 1: Summary of the advantages of the new approach to ... in both screens, with the growth screen having a 1% false discovery rate and identifying >88% of previously...in Morgens et al. contains three types of guides: 1) targeting guides (blue), which target all protein-coding...off-target sites was profiled. Guides with exact or 1-bp mismatch off-targets had greater toxicity than ...characteristics that influence gRNA off-target activity: 1) mismatches closer to the PAM are less tolerated than.... When comparing the toxicity of guides that have 1-bp mismatch off-target sites, short guides had reduced...Knockout Library are available from Addgene! References 1. Morgens, David W., et al. "Genome-scale measurement...
Showing: 101 - 110 of 605 results