Viral Vectors & Particles

GCaMP is a genetically encoded calcium indicator (GECI) and created from a fusion of green fluorescent protein (GFP), calmodulin (CaM), and M13, a peptide sequence from myosin light chain kinase. When GECI binds Ca2+, the conformational change of GCaMP induces proximity of the GFP parts, eliciting strong GFP fluorescence signal, which allows measuring action potentials and other receptor activation events that trigger Ca2+ influx. The advantage of GECI's are that they can be genetically specified for studies in living organisms. GCaMP6 is a novel ultra-sensitive format of GCaMP that outperformed other sensors in terms of accuracy and sensitivity in measuring cytosol Ca2+ level. Mutations in calmodulin part of GCaMP6 created several variants which showed different calcium fluorescence decay rate (fast - GCamP6f, slow - GCaMP6f and medium - GCaMP6m).

Adeno-Associated Virus-DJ (AAV-DJ) is a synthetic serotype made from eight different wild-type AAV serotypes (AAV2, 4, 5, 8, 9, avian, bovine, and goat AAV) using DNA shuffling. These modifications give the AAV-DJ serotype improved transduction efficiency in vitro and in vivo compared to wild-type serotypes. Consequently, AAV-DJ can infect a broad range of cell types. These AAV particles constitutively express ZsGreen under a CMV promoter. ZsGreen is a human codon-optimized variant of the green fluorescent protein isolated from reef coral (Zoanthus sp). It has been engineered for higher expression in mammalian cells and is up to four times brighter than enhanced GFP (eGFP). ZsGreen expression and AAV transduction efficiency can easily be verified and optimized by fluorescence microscopy or flow cytometry. ZsGreen has an excitation wavelength of 493 nm and an emission wavelength of 505 nm.

Adeno-Associated Virus Serotype 4 (AAV4) contains an expanded p5 promoter region compared to either AAV2 or AAV3. The AAV4 Rep gene product shows greater than 90% homology with the Rep products of AAV2 and AAV3. AAV4 can transduce human, monkey, and rat cells, and efficiently transduces type B astrocytes in the subventricular zone and glia overlying the rostral migratory stream neural tube. These AAV particles constitutively express ZsGreen under a CMV promoter. ZsGreen is a human codon-optimized variant of the green fluorescent protein isolated from reef coral (Zoanthus sp). It has been engineered for higher expression in mammalian cells and is up to four times brighter than enhanced GFP (eGFP). ZsGreen expression and AAV transduction efficiency can easily be verified and optimized by fluorescence microscopy or flow cytometry. ZsGreen has an excitation wavelength of 493 nm and an emission wavelength of 505 nm.

Adeno-Associated Virus-DJ (AAV-DJ) is a synthetic serotype made from eight different wild-type AAV serotypes (AAV2, 4, 5, 8, 9, avian, bovine, and goat AAV) using DNA shuffling.  These modifications give the AAV-DJ serotype improved transduction efficiency in vitro and in vivo compared to wild-type serotypes. Consequently, AAV-DJ can infect a broad range of cell types. These AAV-DJ particles constitutively express the firefly (Photinus pyralis) luciferase under the control of a CMV promoter.

Adeno-Associated Virus-DJ (AAV-DJ) is a synthetic serotype made from eight different wild-type AAV serotypes (AAV2, 4, 5, 8, 9, avian, bovine, and goat AAV) using DNA shuffling. These modifications give the AAV-DJ serotype improved transduction efficiency in vitro and in vivo compared to wild-type serotypes. Consequently, AAV-DJ can infect a broad range of cell types.<u> </u> These AAV particles constitutively express the firefly (Photinus pyralis) luciferase and enhanced Green Fluorescent Protein (eGFP) genes connected via a T2A linker, under the control of a CMV promoter. The T2A self-cleaving peptide (derived from Thosea asigna virus 2A) leads to the efficient cleavage of the transcript, and expression of luciferase and eGFP as two separate proteins.

Adeno-Associated Virus-DJ (AAV-DJ) is a synthetic serotype made from eight different wild-type AAV serotypes (AAV2, 4, 5, 8, 9, avian, bovine, and goat AAV) using DNA shuffling. These modifications give the AAV-DJ serotype improved transduction efficiency in vitro and in vivo compared to wild-type serotypes. Consequently, AAV-DJ can infect a broad range of cell types. These AAV particles constitutively express the firefly (Photinus pyralis) luciferase and mCherry genes connected via a T2A linker, under the control of a CMV promoter. The T2A self-cleaving peptide (derived from Thosea asigna virus 2A) leads to the efficient cleavage of the transcript, and expression of luciferase and mCherry as two separate proteins.

Cas9 is an endonuclease enzyme that introduces a double stranded break into the DNA when recruited to a specific DNA sequence by the sgRNA (single guide RNA). This double stranded break is repaired in mammalian cells either through Non-Homologous End Joining or Homologous Recombination. Non-Homologous End Joining often results in the deletion or insertion of several base pairs at the cut site, which, when resulting in a frameshift, causes the functional inactivation of the gene. SaCas9 (Staphylococcus aureus CRISPR-associated protein 9) has demonstrated high cutting efficiency in mammalian cells, and its smaller size makes it ideal for packaging into AAV. SaCas9 recognizes a longer protospacer adjacent motif (PAM) site, 5'-NNGRRT-3', than the more traditional SpCas9 (Streptococcus pyogenes CRISPR-associated protein 9). These AAV particles constitutively express SaCas9 under the control of a CMV promoter. Adeno-Associated Virus-DJ (AAV-DJ) is a synthetic serotype made from eight diffe