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Addgene
Showing: 41 - 60 of 140 results
  1. Google Forums Round Up: First Impressions of NgAgo

    Type
    Blog Post
    ...NgAgo First, unlike Cas9, NgAgo does not require a PAM sequence for target recognition, which gives a researcher...multiple sites in a single experiment. Also, the NGG PAM sequence might not always be available at an optimal...
  2. CRISPR 101: Homology Directed Repair

    Type
    Blog Post
    ...Cas enzymes can continue to cleave DNA if the gRNA/PAM site remains intact. In other words, Cas9 will keep...mutations after an HDR event, ensure the gRNA or PAM site is disrupted in the donor. Choosing a type ...
  3. What's New in CRISPR - May 2019

    Type
    Blog Post
    ... Nme2Cas9, that recognizes a simple dinucleotide PAM (N4CC), lending to high target site density (average...
  4. Zhang Lab's CRISPR Frequently Asked Questions

    Type
    Collection
    ...There is no need to add the NGG PAM sequence. Because the 'NGG' of the PAM is used to select your genomic...target site sequence (protospacer+PAM) is located? If there are other PAMs in the region, will they also ... the PAM (my target sequence) in my HR template an issue? Yes. If you have intact protospacer+PAM sequence...good way is to mutate the PAM 'NGG' sequence in the HR template. Since the PAM sequence is required for... this Nature Protocol article . Should I add the PAM sequence to the oligo when cloning my target spacers...be targeted? The Cas9 cuts 3-4bp upstream of the PAM sequence. There can be some off-target DSBs using... (gRNA). These considerations only matter if the PAM sequence is immediately adjacent to the nearly-homologous...
  5. CRISPR 101: Off-Target Effects

    Type
    Blog Post
    ...often be proximal to a PAM site for DNA cutting to occur. The RNA sequence and PAM site confer specificity...
  6. Church Lab CRISPR Plasmids

    Type
    Collection
    ...Protospacer-adjacent motifs (PAMs): Please note that NM, ST1, and TD recognize different PAMs than does the classical...experimentally determined these PAMs using a highly stringent selection, which revealed that PAM recognition is more...recognize sequences with PAMs matching: NNNNGA, NNNNGTT, NNNNGNNT, just as well as the PAM determined by bioinformatics... only if the correct protospacer-adjacent motif (PAM) is present at the 3′ end (see Figure). Mammalian...protospacers interact nonlinearly with different PAM sequences to determine overall activity at the site...
  7. CRISPR Guide

    Type
    Collection
    ...adjacent to a P rotospacer A djacent M otif (PAM). The PAM sequence serves as a binding signal for Cas9...found in the REC2, REC3, and PAM interacting domains and allows for expanded PAM recognition as well as increased... non-NGG PAM sequences . Another Cas9 alternative is xCas9 , which targets a broad set of PAM sequences...identifying all PAM sequences within the genetic region to be targeted. If there are no PAM sequences for...Cas enzymes ( see Cas9 variants and PAM sequences ). Once possible PAM sequences and putative target sites... near the PAM sequence, so gRNAs with no homology or those with mismatches close to the PAM sequence will...of Cas Enzymes Cas Enzymes and PAM Sequences Species/Variant of Cas9 PAM Sequence* Streptococcus pyogenes...
  8. CRISPR Plasmids - gRNAs

    Type
    Collection
    ... may only be compatible with a single species or PAM binding variant of Cas9. For instance, wild-type ...used with targets that are upstream of a 5' NGG 3' PAM sequence. Which CRISPR application is this gRNA sequence...systems with wiki entries describing enzyme activity, PAM requirements, experimental considerations, and more...
  9. CRISPR History and Development for Genome Engineering

    Type
    Collection
    ...with alternative PAM sites . Cpf1 , another CRISPR nuclease, recognizes a 5’-TTN PAM, improving targeting... sequence known as a protospacer adjacent motif (PAM). The CRISPR complex binds to the foreign DNA and...it contains a special Protospacer Adjacent Motif (PAM) directly after where the crRNA would bind. (5) After...stranded DNA target and cleaves both strands near the PAM. (6) The crRNA:tracrRNA:Cas9 complex unbinds after...bacterial genomes, are highly diverse, with varying PAM sequences and number/types of Cas proteins. Makarova...packaged in AAV. Cas9 orthologs also have distinct PAM sites that do not cross react, allowing multiple ...2015. Engineered CRISPR-Cas9 nucleases with altered PAM specificities. Nature . 523(7561):481-5. PMID: 26098369...
  10. Joung Lab CRISPR-Cas/RNA-Guided Nuclease (RGN) Plasmids

    Type
    Collection
    ...variants with no detectable off-target effects Modified PAM specifity of Staphylococcus aureus Cas9 (SaCas9 KKH...variant) Engineered CRISPR-Cas9 nucleases with altered PAM specificities Dimeric CRISPR RNA-guided FokI Nucleases...
Showing: 41 - 60 of 140 results