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We narrowed to 72 results for: AAV Cas9

Showing: 21 - 40 of 72 results

  1. 15 Hot Plasmids from 2017

    Type
    Blog Post
    Published
    Jan. 9, 2018, 3:02 p.m.
    ..., a base editor containing high fidelity Cas9 variant HF-Cas9. They subsequently tested the original BE3...-BE3  EQR-Cas9 (NGAG)  Lower off-target activity than BE3  85173  pBK-VRER-BE3  VRER-Cas9 (NGCG)   ...The popular CRISPR base editing technique uses dCas9/Cas9 nickase fused to a cytidine deaminase for targeted...Plasmid ID Plasmid Name Cas9 Variant Speacial Features 85169  pJL-SaBE3  SaCas9 (NNGRRT)    85170  pJL-SaKKH-BE3... pJL-SaKKH-BE3  SaCas9 (NNGRRT)    85171  pBK-VQR-BE3  VQR-Cas9 (NGA)  Lower off-target activity than...the effects of dCas9 binding to a genomic locus, or limiting the amount of time that Cas9 is active in ...interneurons, Gordon Fishell’s lab has developed a new AAV approach using regulatory elements that restrict ...

  2. Hot Plasmids - May 2022

    Type
    Blog Post
    Published
    May 26, 2022, 1:15 p.m.
    ... of the AAV capsids (left) and transduction patterns (right).  Both AAV-PHP.eB and the AAV.CAP-B capsids... with Cas9 editors. The lab believes that this system could potentially be packaged into an AAV system...expression using Cre-mediated AAV Targeted Evolution method (M-CREATE) dCas9-SSAP with reduced error rate...Increased neuronal expression using Cre-mediated AAV Targeted Evolution method (M-CREATE) by: Brian O'...developed the multiplexed version of the Cre-mediated AAV Targeted Evolution method (M-CREATE), in which one...of up to 20%, comparable to commonly used CRISPR/Cas9 methods for large inserts. However, this system ...inactive dCas9 and microbial SSAPs (dCas9-SSAP) which insert genes in a cleavage free manner. The dCas9-SSAP...

  3. Viral Vectors 101: Viral Applications

    Type
    Blog Post
    Published
    April 13, 2023, 1:15 p.m.
    ...specifically AAV, are also being exploited as delivery mechanisms for gene therapy. The tractability of AAV, low...Cepko Discuss Gene Therapy with AAV Adenoviral Delivery of CRISPR/Cas9 Aims to Expand Genome Editing to...of virus to choose from (lentivirus, retrovirus, AAV, etc.). When you’ve decided on an application, our...depending on the vector and virus. Viral CRISPR-Cas9 tools have been engineered to introduce frameshift...00487-6 Additional Resources on the Addgene Blog AAV: A Versatile Viral tool for Gene Expression in Mammals...

  4. AAV Q&A with Tim Miles

    Type
    Blog Post
    Published
    May 17, 2022, 1:15 p.m.
    ...questions!  AAV Prep and Technique Q1: Is there any way to deliver a long gene (>4.5 kB) by AAV? A: One potential... make an efficient AAV prep for a 4.2 kB insert with the CMV promoter using the AAV-BR1 capsid? A: It ...our understanding that AAV-BR1 has the same genome size restrictions as other AAV. Therefore, one should...samples within a project. Targeting Your AAV Q4: Are there any AAVs that have successful transduction in ...beginners Adenoviral delivery of CRISPR/Cas9 Tips for a first time AAV user Resources on Addgene.org Find...animal models. All of our engineered AAV are presently built on an AAV9 backbone and so will very likely ...Did you catch our April AAV webinar with Tim Miles, PhD, Director of the CLOVER Center at CalTech? If...

  5. Hot Plasmids and Viral Preps - January 2021

    Type
    Blog Post
    Published
    Jan. 5, 2021, 2:15 p.m.
    ... AAV8, as well as AAV5 and AAVrg. Find the Cre-expressing viral preps here AAV-CAG-dLight1.3b from ...current N-terminal linked SaCas9 ABE variants. It's also one of the smallest AAV-deliverable ABE.  Find...in serotype AAV5! Get dLight1.3b here Looking for a knock-out in your experiments? AAV-flex-taCasp3...available as a ready to use viral prep in serotype AAV5. Find AAV-flex-taCasp3-TEVp here The red-shifted channelrhodopsin... ~70kDa, with a molecular weight roughly half of Cas9 or Cas12a. This is in part due to the fact that ...CRISPR Plasmids and Resources page.  CDDR (CRISPR–Cas9-based Dual-fluorescent DSB Repair) is a double-strand...the Deisseroth viral preps here Coming soon! New AAV serotypes from the Sanford Boye lab with enhanced...

  6. Editor's Choice, July 2016

    Type
    Blog Post
    Published
    Aug. 5, 2016, 3 p.m.
    ...Generating Mouse Models Using CRISPR/Cas9 Cpf1 Update: Comparison to Cas9 and NgAgo Special Delivery: Fluorophore... when compared to the most commonly used Cas9 variant, SpCas9. These properties, combined with other benefits...generate mouse models.   Cpf1 Update: Comparison to Cas9 and NgAgo As you might expect given the excitement...Gendered Innovations: Why Does Sex of the Cell Matter? AAV: A Versatile Viral Tool for Gene Expression in Mammals...

  7. Comparing Cas9 to NgAgo: Can the Argonautes Best CRISPR?

    Type
    Blog Post
    Published
    June 9, 2016, 2:30 p.m.
    ...lines, with an efficiency similar to that of Cas9. Like Cas9, NgAgo is also able to mediate homology-directed...at Addgene Cas9 and NgAgo head to head NgAgo has a number of unique features compared to Cas9. These include...the Addgene Blog Read About Cas9 Homolog Cpf1 Check Out High Fidelty Cas9 Mutants Browse All Genome Engineering... why you might choose NgAgo over CRISPR proteins Cas9 or Cpf1 - keep reading to learn more! Background...include: More targeting flexibility Unlike Cas9 and the other CRISPR enzymes, NgAgo does not require a...genomic target. NgAgo also appears to outperform Cas9 in GC-rich regions. DNA guides NgAgo uses 5’ phosphorylated... “gDNAs”) rather than the RNA guides employed by Cas9 or Cpf1. These short, 24 base DNA guides may be ...

  8. Hot Plasmids - October 2020

    Type
    Blog Post
    Published
    Oct. 13, 2020, 1:15 p.m.
    ... as AAV5. Find the AAV prep here The red-shifted calcium sensor jRGECO1a is now available as AAV-PHPeb...mRNA display (FingRs), Xue Han’s lab has developed AAV vectors to selectively visualize excitatory and inhibitory... codon-optimized, engineered Streptococcus canis Cas9 nuclease that has a broad 5′-NNG-3′ PAM. Get the...Applications (CHyMErA) Knockout Libraries express hybrid Cas9-Cas12a gRNAs under a single U6 promoter. The paralog...AAV-PHPeb. Find the AAV prep here ...

  9. What's New in CRISPR - September 2019

    Type
    Blog Post
    Published
    Sept. 10, 2019, 12:56 p.m.
    ...inducible CRISPR/Cas9 systems for transcriptional activation and genome editing by fusing Cas9 to a mutated...vectors to create knock-ins CasX Drug inducible CRISPR/Cas9 activation E. coli genome-wide inhibition library...vectors GeneWeld from Jeffrey Essner’s lab uses CRISPR/Cas9 and a donor vector to create knock-ins of fluorescent..., or transcriptional regulators. In this system, Cas9 generates a double-stranded DNA break at the genomic...Nucleic Acids Research Find the drug inducible Cas9 plasmids E. coli genome-wide inhibition library...factors, and identify sequence-specific toxicity of Cas9. Read the paper in Nature Communications Find...Tumor Suppressor Gene CRISPR Knockout Library is an AAV CRISPR knockout library targeting mouse tumor suppressor...

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

    Type
    Blog Post
    Published
    May 3, 2018, 1:48 p.m.
    ...smaller than Cas9! Their small size makes then easy to package in low-capacity vectors like AAV, a popular...significant off-targets, CasRx displayed zero. Unlike Cas9, for which efficiency varies widely across guide...acceptor and two putative splice enhancers. After AAV-mediated delivery to iPS-derived cortical neurons...CRISPR RNA Editing with REPAIR Get tips for using Cas9 Transcriptional Activators Learn about Cas13a (C2c2...regulating methods: small hairpin RNA interference, dCas9-mediated transcriptional inhibition (CRISPRi), and...

  11. Using CRISPR/Cas9 to Edit Disease Out of the Genome

    Type
    Blog Post
    Published
    June 23, 2022, 1:15 p.m.
    ...Northeastern University.  There can be no doubt that CRISPR/Cas9 technology has been a breakthrough for the genome-editing...lab are researching a way to treat AD using CRISPR/Cas9. They are targeting the genes responsible for both... AD. In this study, the group showed that CRISPR/Cas9 could be used to edit several autosomal-dominant...that play into late onset AD (5).     CRISPR/Cas9 corrects Alzheimer’s disease mutation in basal forebrain... could be rescued by coinjection into zygotes of Cas9 mRNA and a single-guide RNA targeting the mutant...Hubrecht Institute in The Netherlands used the CRISPR/Cas9 genome editing system to correct the cystic fibrosis...cystic fibrosis (8).   The difference with CRISPR/Cas9 in comparison to other genome editing techniques...

  12. CRISPR 101: Cas9 vs. The Other Cas(s)

    Type
    Blog Post
    Published
    Dec. 8, 2022, 2:15 p.m.
    ...similarities and differences to Cas9. DNA targeting Cas enzymes Cas9 – The genome engineering workhorse...CRISPR’ without shortly thereafter hearing ‘Cas9’. The Cas9 nuclease is a genome engineering powerhouse...making Cas9 an adaptable choice for many genome engineering endeavors. For more information on Cas9 variants...same mechanism as traditional spCas9. If you are looking to modify RNA, Cas9 just won’t cut it – literally...We are all oh so familiar with the nuclease Cas9, but how does it stack up to the new Cas enzymes on ...proceed to generate a blunt DNA double strand break. Cas9 has been studied for genome engineering purposes...variants, check out our blog post on choosing a Cas9 enzyme. Fast facts – PAM requirement: NGG and an expanding...

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

    Type
    Blog Post
    Published
    Nov. 19, 2020, 2:15 p.m.
    ...the researchers developed adeno-associated viral (AAV) vectors for the targeted delivery of a functional...MECP2 protein, showing mutations corrected by CRISPR/Cas9 in wild-type human iPSCs. Image modified from Le...Le et al., 2019. Gene editing via CRISPR/Cas9 and RNA-mediated correction of MECP2 mutant alleles,... Klaus Rajewsky’s group recently used the CRISPR/Cas9 system to both precisely insert the MECP2-R270X ...Joung, PhD, and Feng Zhang, PhD, is pursuing CRISPR/Cas9 technology for precise correction of Rett syndrome...Syndrome mouse model using an adeno-associated viral (AAV) vector. The editase repaired half of the mutant ...Rajewsky K, Chu VT (2019) Efficient and Precise CRISPR/Cas9-Mediated MECP2 Modifications in Human-Induced Pluripotent...

  14. Four Ways to Package Transgenes That Exceed the Size Limit of Adeno-associated Virus

    Type
    Blog Post
    Published
    Sept. 1, 2020, 1:15 p.m.
    ...time AAV user Learn about how AAV is used in retinal gene therapy Learn about the parts of an AAV transfer... of ~9 kb into AAV. They found that in defiance of the packaging limit dogma, this AAV successfully mediated...size of the AAV donor template for CRISPR-mediated homology directed repairs (HDR). Split AAV vector approaches...independently packaged into an AAV. When a cell is co-infected with both part A and part B AAVs, the fragments reassemble...Addgene.org: Check out the Addgene AAV guide Find ready-to-use AAV preps from Addgene's viral service ...Adeno-associated virus (AAV) has many features which make it a great viral vector, but its packaging ...how do you fit a big gene into a tiny vector like AAV? By breaking the transgene into smaller pieces. Reassembly...

  15. Adenoviral Delivery of CRISPR/Cas9 Aims to Expand Genome Editing to Primary Cells

    Type
    Blog Post
    Published
    Dec. 10, 2020, 1 p.m.
    ...capacity larger than AAV vectors. This capacity is large enough to carry the Cas9 gene and a gRNA expression...DNA Adenoviral CRISPR/Cas9 genome editing tools can also be used to deliver Cas9 and DNA templates in ... CRISPR/Cas9 research! To find more information about the adenoviral delivery of CRISPRS/Cas9 using the...Genome Editing of Pten by CRISPR/Cas9 in Mouse Liver in Spite of Cas9-Specific Immune Responses. Human... out the plasmids at Addgene: pAdSh.PGK.Cas9 (expresses S. pyogenes Cas9 from the PGK promoter) and U6...left) and targeted DNA knock-in (right) by CRISPR/Cas9. Adenoviral CRISPR vectors (left) or CRISPR vectors...transduced to target cells. Within target cells, Cas9 machinery is transiently expressed and binds to ...

  16. Cpf1: A New Tool for CRISPR Genome Editing

    Type
    Blog Post
    Published
    Oct. 14, 2015, 2:30 p.m.
    ...needed by Cas9 for cleavage. In certain cases, Cpf1 may be better suited for genome editing than Cas9 - read...CRISPR system. Like Cas9, Cpf1 contains a RuvC-like endonuclease domain, but it lacks Cas9’s other HNH endonuclease...Potential advantages of Cpf1 over Cas9 Type II CRISPR systems based on Cas9 were thought to be the simplest...than that of the widely used SpCas9 and comparable to SaCas9. As seen with Cas9 orthologs, Cpf1 efficiency...rate is similar to that of high fidelity Cas9s eSpCas9 and SpCas9-HF1. Both AsCpf1 and LbCpf1 ribonucleoproteins...needed for Cas9 function. Since both Cpf1 and its guide RNAs are smaller than their SpCas9 counterparts...display cleavage activity in mammalian cells. Like Cas9 nucleases, Cpf1 family members contain a RuvC-like...

  17. DAP arrays for multiplex gene editing

    Type
    Blog Post
    Published
    July 7, 2022, 1:15 p.m.
    ... found led them back towards Cas9 fused base editors (nCas9-BE), as Cas9 is mechanistically more favored...also exhibited reduced Cas9-dependent off-target editing and do not cause higher Cas9-independent off-target...leading the edge of the precision gene editing. CRISPR-Cas9, the key component to enable efficient base editing...organisms and has previously been used for multiplex Cas9-nuclease editing. Typically, a single array encoding...elements for deliveries via adeno-associated virus (AAV) and lentivirus, demonstrating simultaneous editing...would independently generate multiple gRNAs for nCas9-BE, or any other kinds of CRISPR tools, without ...

  18. CRISPR 101: Anti-CRISPR Proteins Switch Off CRISPR-Cas Systems

    Type
    Blog Post
    Published
    July 23, 2020, 1:20 p.m.
    ...et al., 2016). While Streptococcus pyogenes Cas9 (SpyCas9) is the most used and well studied CRISPR system... study, the labs found that anti-CRISPR prevents Cas9 activity in HEK293 by reducing editing from ~30%...interact with CRISPR-Cas systems such as Cascade-Cas3, Cas9, Cas12, and Cas13 (Marino et al., 2020). Some Acr...ways: Inhibit DNA binding Example:  AcrIIA4 blocks Cas9’s interaction with the PAM site (Dong et al., 2017... crRNA loadingExample: AcrIIC2 interactions with Cas9 interferes and prevents the correct assembly of ... AcrIIC1 binds to the HNH endonuclease domain of Cas9 and prevents target DNA cleavage (Harrington et ... Doudna lab found that at least 50% of on-target Cas9 edits happens within the first six hours. When added...

  19. 10 Great Guest Posts We're Thankful For

    Type
    Blog Post
    Published
    Nov. 18, 2016, 3 p.m.
    ...action. AAV: A Versatile Viral Tool for Gene Expression in Mammals Adeno-associated viruses (AAVs) are commonly...Cepko’s Lab at Harvard Medical School describes how AAVs function and indicates important things to consider... editing - all influenced by an understanding of Cas9 biophysics! Casilio: An Adaptive, Multitasking “... system that uses modular RNA binding proteins, dCas9, and a modified gRNA to direct a variety of customizable...

  20. Meet the New Addgene Board Members!

    Type
    Blog Post
    Published
    April 28, 2020, 1:14 p.m.
    ...project to develop an AAV-based COVID-19 vaccine.  Vandenberghe’s work with AAVs led to many interactions...Therapeutics and is an Addgene depositor. KSQ uses a CRISPR-Cas9 platform to systematically test the function of ...was compelled to be part of that.” He’s deposited AAV constructs for gene delivery. In 2018, he interviewed...
Showing: 21 - 40 of 72 results