Skip to main content
Addgene

Enter a sequence to perform a BLAST-based search of Addgene plasmid sequence databases.

In Category

We narrowed to 567 results for: PAC;

Showing: 61 - 80 of 567 results

  1. Bringing Sustainable Practices to the Lab: Recycling

    Type
    Blog Post
    ...Service picks up mail. Recycle plastic packaging Packaging is a huge contributor to the increase in ...lot of their plastic packaging (from Corning®, Falcon®, or Axygen® product packaging) for free! It has to...the lab could make a big difference in our global impact as scientists. Many scientists are already leading...toward responsibility in our lab’s environmental impact, but we can all do more to make research more environmentally...your address sticker off the box, remove the ice pack (it’s not recyclable), tape the box shut, and drop... to be from their company and it has to be packaging (no coated tissue culture plates, for example). Check...and companies have the potential to make a huge impact directly on waste, but also on culture. Encourage...

  2. Tips for a 1st time AAV user (by a Rookie AAV user)

    Type
    Blog Post
    ...analyses needed for packaging AAV. AAV production Overview: The first step in packaging AAV is transfecting...had never worked with AAV, so naturally I had to package some virus for my first experiment. It was a bit...transfecting HEK293 cells with AAV packaging plasmids. The cells need three different plasmids to produce AAV...intermediate into the single-stranded genome that's then packaged into AAV’s protein shell, or capsid; 2) the pHelper...plasmid which encodes a transgene of interest that’s packaged into the virus. Two to five days after transfection...plasmids contain ITRs which are required for proper packaging of AAV, but also form secondary structures that...spreadsheet similar to this one to plan my AAV packaging PEI transfections. It saves me time and helps ...

  4. Western Blot

    Type
    Protocol
    ...gel as follows: Remove the gel from the plastic packaging. Remove tape and plastic comb. Rinse the wells...

  5. Cas13d: Small RNA-targeting CRISPR enzymes for transcriptome engineering

    Type
    Blog Post
    ...average only 2.8 kb in size and are easy to package in low-capacity vectors! These small, but mighty type VI-D...Cas9! Their small size makes then easy to package in low-capacity vectors like AAV, a popular vector due ...HEPN-independent. These enzymes also do not require a protospacer flanking sequence, so you can target virtually...either an unprocessed or processed gRNA array (22 nt spacer with 30 nt direct repeat) mediating potent knockdown...dementia-associated tau, delivering dCasRx and a 3-spacer array targeting the MAPT exon 10 splice acceptor...toolbox will be. It is, however, certain that the impact of CRISPR scientific sharing will continue to grow...Addgene. Yan, Winston X., et al. “Cas13d Is a Compact RNA-Targeting Type VI CRISPR Effector Positively...

  6. The CRISPR Software Matchmaker: A New Tool for Choosing the Best CRISPR Software for Your Needs

    Type
    Blog Post
    ...think we are currently at in the CRISPR software space. The CRISPR research community hasn't had a chance...review process has been challenged with many closely spaced CRISPR software submissions. With few prior publications...GUI”. Trends: Online tools dominate the software space but generally rely on less powerful algorithms to...user in different formats. This can have a large impact on downstream results. Examples: “HTML”, “visual...agnostic tools are the better solution but are often packaged as offline tools requiring expertise in one programming...are important to define for the CRISPR software space. It is unlikely that you will find these terms anywhere... data dump of ALL strings in a genome using any IUPAC encoded pattern at any edit distance. It won’t, ...

  7. Quick Guide to All Things Lentivirus

    Type
    Blog Post
    ...insert into a viral particle (the packaging signal Psi). Because the packaging plasmids lacked the psi signal...plasmid and the packaging plasmid caused the Psi sequence to be placed on the packaging plasmid forming...virus (MoMLV) and consisted of two plasmids: the packaging plasmid containing all the structural genes necessary...virus. Indeed, by testing several vectors and packaging cell line combinations, Miller et al. showed that...generated by the transfer of a retrovirus into the packaging cell line PA12 (containing the psi deletion). ...viral genes with the exception of the env gene (packaging plasmid), and 3) the vesicular stomatitis virus...293T cells, with your transfer plasmid and your packaging plasmids. For this, you can use the transfection...

  8. The AAVantages of AAV in CRISPR Screens

    Type
    Blog Post
    ...to using AAV is the limited packaging capacity. Lentiviruses have capacities of 8–12 kb, while AAV only...libraries available at Addgene. The large packaging capacity and stable expression of lentiviral vectors...Lentivirus has limited packaging options, with most lentiviral vectors being packaged using VSV-G. In contrast...contrast, AAV can be packaged in a variety of capsids. To date, at least 12 different capsids, or serotypes...that when using Cas9, you’re already almost out of space! Don’t let this get you down for too long, though...

  9. Viral Vectors 101: Integrase-Deficient Lentiviral Vectors

    Type
    Blog Post
    ...further! You can package your own lentiviral vector using an integrase-deficient packaging plasmid available... the integrase-deficient packaging plasmid instead of the standard packaging plasmid. For more details...integration of this portion into the host genome. The packaging plasmid contains Gag, Pol, Rev, and Tat genes....transcriptase), and IN (integrase). These viral packaging genes are required for the production of lentiviral...integrated into the host genome via the viral proteins packaged during production (read our blog post on the retroviral...integrated portion does not contain any of the viral packaging genes, the natural cycle of the lentiviral vector...details on how to package lentiviral vectors, check out Addgene’s Lentiviral Guide and our lentiviral production...

  10. Hot Plasmids: Summer 2024

    Type
    Blog Post
    ...evolved optimizations for prime editing CHARM: A compact epigenetic silencer Recombinant antibodies for ...assembly of biotinylated SpyTag-SpyCatcher linkers and spacer proteins ending with a GFP nanobody (Figure 1)....determination. A) Schematic of workflow. B) Detail of spacer peptides (3HB: 11-nm 3-helix bundle; SAH: 60-nm...software (green circles), magnetic bead (*), and spacers are labeled; scale bar: 100 nm. D) 3D model of ...other targets. The modular system of linkers and spacers could enable many future variations using nanobodies... the PE components to be delivered in a single package. In a recent publication, the Liu lab showed that...doi.org/10.1038/s41551-024-01227-1   CHARM: A compact epigenetic silencer By Emily P. Bentley The Weissman...

  11. Learning Technical Skills at a Lab Central Workshop

    Type
    Blog Post
    ...new teaching space at Lab Central to learn all about it. LabCentral is a lab and office space shared by ...and making space for the techniques ourselves. As you probably have limited time and space in your own...by numerous emerging biotech companies. The space was modern with clean bold colors of orange and white...a few goodies which included an NEB drawstring backpack and a double wall insulated glass to take home...like workshops at Lab Central. Because of the fast pace of work in the Addgene lab, we don’t often get the...opportunity to explore new techniques in our own space. Workshops such as this give us a chance to learn...

  12. Components of CRISPR/Cas9

    Type
    Blog Post
    ...RNA provided that target is upstream (5') of a protospacer adjacent motif (PAM) Cas9 endonuclease binding... RNA and a 3-base pair sequence known as the Protospacer Adjacent Motif or PAM. In order for dsDNA to ... and the unique stretches of DNA between them “spacers” (see figure below). After years of research, we... each direct repeat, combined with its adjacent spacer, ultimately encodes a single crRNA. The direct ...pre-crRNA into mature crRNA and tracrRNA binding. The spacer regions, on the other hand, are the unique, foreign...tracrRNA forms the scaffold portion of a gRNA and the “spacer region” forms the target sequence.   Overview...transcribed into pre-crRNAs containing both the spacer region and the direct repeat region. B) RNase III...

  13. CRISPR History and Development for Genome Engineering

    Type
    Collection
    ...contains many unique protospacer sequences that have homology to foreign DNA. Protospacers are separated by...Adaptive Immune System CRISPR (Clustered Regularly Interspaced Short Palindromic Repeat) sequences were initially...contain a species-specific sequence known as a protospacer adjacent motif (PAM). The CRISPR complex binds...sequence is only valid if it contains a special Protospacer Adjacent Motif (PAM) directly after where the... development has occurred at a truly astonishing pace, with work directed towards increasing on-target...fidelity Cas9 variants , SpCas9-HF, eSpCas9, and HypaCas9 display very low off-target activity due to rationally...SaCas9) is smaller than SpCas9 and more easily packaged in AAV. Cas9 orthologs also have distinct PAM ...

  14. Back to Bacteria: CRISPR gRNA Multiplexing Using tRNAs

    Type
    Blog Post
    ...Cas9 multiplexing in systems with limited packaging capacity. Still confused about the various tRNA expression... will be necessary to improve its multiplexing capacity. Multiplexing is key due to the natural redundancy...often necessary. Guide RNAs (gRNAs) are commonly packaged in 400-500 bp cassettes containing the RNA pol...100 bases) limit the number of gRNAs that can be packaged together in a single vector. In addition, the ...forms tRNA-gRNA and tRNA-gRNA-tRNA, with short 5’ spacer sequences preceding gRNAs, to test for gRNA cleavage...predicted with no addition of nucleotides to the 5’ spacers. The 3’ ends were modified slightly with either...once, and each <200 bp tRNA-gRNA takes up half the space of a standard pol III-gRNA, which will facilitate...

  15. Delivery Methods for Generating iPSCs

    Type
    Blog Post
    ...nondividing cells and have an ~8kb packaging capacity. With this packaging capacity, reprogramming factors can...other delivery methods. Retrovirus has a cloning capacity of ~8 kb, so either a single polycistronic vector...or multiple single-gene vectors can be used to package reprogramming factors. An important milestone of...and nondividing cells and have a larger cloning capacity (8-10 kb) than other retroviral vectors. This ...a vector and chromosomal DNA. PB has a cloning capacity of ~9-14 kb. This system is transfected into cells...simple and fast to evaluate. iPSCs should form compact colonies that are comprised of cells with a large...differentiated in vitro to form embryoid bodies which are compact balls of loosely organized tissues that express...

  16. What's New in CRISPR - November 2021

    Type
    Blog Post
    ...system is that it is too large to be effectively packaged into AAV particles for delivery into mammalian... much smaller and would be ideal for such AAV packaging, but their activity in mammalian systems has been... based systems, making it well suited for AAV packaging. Most importantly, this CasMINI system was shown...CasMINI plasmids Efficient CRISPR editing with a hypercompact Cas12f1 and engineered guide RNAs Yong-Sam Kim...result is a highly efficient Cas system with very compact genome editors and a gRNA down-sized by almost ...specifically binds its own mRNA to facilitate its packaging into virus-like particles. Feng Zhang’s lab adapted...the native PEG10 RNA sequences to instead drive packaging of cargo sequences in the capsid, in an approach...

  17. Kit Free RNA Extraction

    Type
    Protocol
    ...hazards for these reagents. Work in a well-ventilated space and under a fume hood when working with the volatile...

  18. The Effect of COVID-19 on Liu Lab Plasmid Requests From Addgene Data

    Type
    Blog Post
    ... across regions, with Europe most impacted and China least impacted. The inferred weekend effect is, in...date range is, they could be the most impacted or least impacted. These results highlight the instability... last six months, COVID-19 has had a tremendous impact on our world -- as of June 4, 2020, COVID-19 has...unique PIs/labs placing orders per day) on dates impacted by each effect. The inferred parameters suggest...methods for fundamental scientific discovery and high-impact applications.       Alvin Hsu is a graduate student...next-generation therapeutics. Prime editing, base editing, PACE, and DNA-templated synthesis are four examples of...

  19. Genomic Deletions in Mammalian Cell Lines

    Type
    Collection
    ...consist of a 20-mer (“protospacer sequence”) upstream of an “NGG” sequence (“protospacer adjacent motif” or...Abstract The prokaryotic clustered regularly interspaced short palindromic repeats (CRISPR)/CRISPR-associated...Figure 2A ). NOTE: In this example, sgRNA-A’s protospacer sequence and PAM happen to fall on the bottom...bottom (Crick) strand while sgRNA-B’s protospacer sequence and PAM fall on the top (Watson) strand ( Figure...will occur independent of orientation of the protospacer sequence/PAM relative to the top or bottom strand...However, if the first position of the 20-mer (protospacer sequence) is G, do not add another G ( e.g., ...cleavage site to ensure detection would not be impacted by a small indel at the sgRNA target site. Design...

  20. Viral Genome Sequencing - A One Stop Shop for AAV Identity and Serotype Confirmation

    Type
    Blog Post
    ...amount of spurious DNA from the packaging cell line, helper and packaging plasmids. We decided to use this... genome exists as [+] and [-] strands that are packaged at the same frequency, we reasoned that, following...viral genome sequencing to determine serotype AAV packaging is not perfect and while we think of the AAV vector...not require any additional reagents or lab work (Pacouret, 2017). Moreover, the sequencing data from multiple...Method for Rapid and Inexpensive Validation of Packaged Single-Stranded Adeno-Associated Viral Genomes...195–205 . https://doi.org/10.1089/hgtb.2019.110 Pacouret S, Bouzelha M, Shelke R, Andres-Mateos E, Xiao...
Showing: 61 - 80 of 567 results