... available with multiple Cas9 variants, including high fidelity Cas9s, Cas9s with various PAM requirements... NAG binding) SpCas9 VRER variant 3' NGCG SpCas9 EQR variant 3' NGAG SpCas9 VQR variant 3' NGAN or NGNG...Species/Variant of Cas9 PAM Sequence* Streptococcus pyogenes (SP); SpCas9 3' NGG SpCas9 D1135E variant 3' ... engineering. Read more about Cas9 variants . Anti-CRISPR Prolonged Cas9 activity can increase the risk...LbCpf1) 5' TTTV AsCpf1 RR variant 5' TYCV LbCpf1 RR variant 5' TYCV AsCpf1 RVR variant 5' TATV Campylobacter...strand SpCas9-HF1 - disrupt Cas9’s interactions with DNA phosphate backbone HypaCas9 - increase Cas9 proofreading...PAM-flexible or PAMless Cas9s. SpCas9-derived variants with non-NGG PAM sequences include: xCas9 - NG, GAA, and...

...organism without a specific preferred mutation Cas9 (or Cas9 nickase) Single (or dual) gRNA targeting 5′ ...generating a point mutation or inserting a tag Cas9 (or Cas9 nickase); Base editor Single (or dual) gRNA ...concern, Cas9-gRNA ribonucleoprotein (RNP) complexes are advantageous due to the transient Cas9 expression... RNA delivery of Cas9 and gRNA In vitro transcription reactions generate mature Cas9 mRNA and gRNA, which...generate transgenic embryos Cas9-gRNA ribonucleoprotein (RNP) complexes Purified Cas9 protein and in vitro transcribed...want to consider alternative Cas enzymes ( see Cas9 variants and PAM sequences ). Once possible PAM sequences...and/or Cas9 expression or (2) a lack of efficient target cleavage in cells expressing both Cas9 and gRNA...

...Uses Viral Integration Serotypes Pseudotyping AAV Variants References AAV Components The small (4.8 kb) ssDNA...plasmids available from Addgene. The popular CRISPR/Cas9 genome editing system has been modified for use ...of AAV-DJ, but with enhanced brain uptake. AAV Variants As mentioned above, AAV improvements have included...hybrid viruses with new properties. Other system variants include: Self-complementary AAV (scAAV): One downside...Mol Ther. 2009 Aug;11(4):442-447. PubMed . AAV variants: Self-complementary recombinant adeno-associated...use in AAV, or containing Staphylococcus aureus (SaCas9) . AAV is also a promising method for gene therapy...