...the use of pAdEasy-2 can increase the capacity of the adenoviral vector. pAdEasy-2 does not contain the...Adenovirus packaging plasmid that lacks E1 and E3 pAdEasy-2 Adenovirus packaging plasmid that lacks E1, E3, and...compatible with the pAdEasy-2 packaging plasmid; Supplies viral protein E4 E1 Supplied in trans by 293...AdEasier-1 cells BJ5183 E. coli cells containing the pAdEasy-1 packaging plasmid 911E4 cells Cell line compatible...then transformed into AdEasier-1 cells , which are BJ5183 E. coli cells containing pAdEasy™. pAdEasy™ is... 293/911 cells throughout virus production. During experiments, GFP can be used to sort cells infected...must be transfected into 911E4 cells for virus production, as 293 cells do not contain E4. Adenovirus ...

...nonspecific cation influx into the cell when stimulated. Channelrhodopsin-2 (ChR2), the first widely adopted... (Phy) interaction 660 Cryptochromes Cryptochrome 2 and CIB1 Blue-light–mediated induction of protein ...interactions based on Arabidopsis thaliana cryptochrome 2 and CIB1 450 LARIAT Inhibits protein function by reversibly...customizable TALE DNA-binding domain with cryptochrome 2 protein and its interacting partner CIB1 to control...transcription effectors OPTOSTIM PHR domain2 of cryptochrome 2 (Cry2) from Arabidopsis thaliana is fused to truncated... with single-spike temporal resolution. PLoS One. 2(3):e299. PMID 17375185 Kim B, Lin MZ. 2013. Optobiology...comparative analysis of microbial opsins. Nat Methods. 9(2):159-72. PMID 22179551 Saunders A, Johnson CA, Sabatini...

...capsids from serotypes 1, 2, and 5 display differential efficiency and cell tropism after delivery to ...indicates a virus containing the genome of serotype 2 packaged in the capsid from serotype 5. Use of these...neurons that are not efficiently transduced by AAV2/2, and is distributed more widely in the brain, indicating...PJ, Mandel RJ, Muzyczka N. Mol. Ther. 2004 Aug;10(2):302-17. PubMed . From virus evolution to vector revolution...persist in non-dividing cells for a period of years without damaging the host cell. In contrast, viral particles...through cell lysis. Episomal stability enables long-term transgene expression in non-dividing cells and is... the helper plasmid are transfected into HEK293 cells, which contain the adenovirus gene E1+, to produce...

...muscarinic receptor and has not been used in vivo . Figure 2: DREADDs, their ligands, and signaling properties.... Reference Rq(R165L) Human M3 muscarinic Arrestin-2/-3 CNO* Increase Arrestin translocation Arrestin signalling...hM1Dq hM5Dq Human M3 muscarinic G αq CNO* Increase Ca 2+ Neuronal burst firing Armbruster et al., 2007 rM3D...channels, their effect, and outcome in neurons. Table 2. Descriptions of PSAMs and their activity in neurons...Forbes SL, Liggett SB (2001). Modification of the beta 2-adrenergic receptor to engineer a receptor-effector...targeted to specific tissues, cell types or even subcellular regions of a neuron. Cell-specific expression of...Human Stem Cell-Derived Neurons Repair Circuits and Restore Neural Function. Cell Stem Cell. 28:112-126...

...Haploid Embryonic Stem Cells Carrying a Guide RNA Library. Cell Stem Cell . 17 (2), 221–232. PMID: 26165924...used to transduce your experimental cell population (Figure 8E). In a 2-vector system, you’ll need to either...of CRISPR-Cas9 with Bacteriophage Proteins. Cell , 168 (1–2), 150-158.e10. PMID: 28041849 Sakuma, T., Nishikawa... regulation of transcription in eukaryotes. Cell . 154 (2), 442–451. PMID: 23849981 Gilbert, L. A., Horlbeck...Transcriptional Programs with CRISPR RNA Scaffolds. Cell . 160 (1–2), 339–350. PMID: 25533786 Cas9 alternatives...single RNA-Guided endonuclease of a Class 2 CRISPR-CAS system. Cell , 163 (3), 759–771. PMID: 26422227 Zetsche...left free to interact with the target DNA. Figure 2: Overview of the NHEJ repair mechanism Cas9 will only...

...surface of most immature B cells; function is unknown Monomer with a valency of 2 IgE Protects against parasites...such as anaphylactic shock Monomer with a valency of 2 IgG The most abundant isotype in blood, also found...secondary immune response Monomer with a valency of 2 IgG1; IgG2; IgG3; IgG4 IgM Produced during the primary... ranging from antibody fragments (shown in Figure 2) to completely unrelated molecules. Examples include...used for assembling protein nanostructures. Figure 2: An antibody (IgG, far left) compared to alternative... produced. Cells - Antibodies can also be produced in cell cultures using hybridomas, cell cultures created...created by fusing B cells with myeloma cells (immortalized B cells). The genetic reorganization that creates...

...specific control of activity. Read More CRISPR Class 2 C lustered R egularly I nterspaced S hort P alindromic...receptors to selectively control and investigate cell signaling pathways. Receptors are remotely controlled...and genetic engineering to measure and manipulate cells (frequently neurons) and their governing biomolecular...detect, measure, and control molecular signals and cells in order to understand their functions. Read More...

... D, Nerhbass U, Montagnier L, Charneau P. Cell. 2000. 101(2): 173-185. PubMed . Woodchuck Hepatitis Virus...selection. Desfarges S, Ciuffi A. Viruses. 2010. Jan;2(1):111-30. PubMed . Altering the tropism of lentiviral...can use FACS to sort cells expressing GFP and later expand these cells into a cell line. Where does lentivirus...actively dividing cell types whereas standard retroviruses can only infect mitotically active cell types. This... necessary. What cell line should be used in order to produce lentivirus? 293T cells are usually used ...be used to make stable cell lines? Lentiviruses can be used to make stable cell lines in the same manner...infected host cells. When these antibiotics are added to the growth medium of the host cells, they kill ...

... or (2) a lack of efficient target cleavage in cells expressing both Cas9 and gRNA. Edited cells may be...expressed in your target cells. For easy-to-transfect cell types (e.g. HEK293 cells), transfection with standard... CRISPR machinery. For more difficult cells (e.g. primary cells), viral delivery of CRISPR reagents may...identify and enrich positive cells, or selection marker to generate stable cell lines (for details, see Plasmids...wide variety of mammalian cell lines Useful for difficult to transfect cell types, can also be used in...possible genotypes within the resulting cell population. Some cells may be wild type due to either (1) a ...will use the example of CRISPR/Cas9 in mammalian cells, many of these principles apply to using CRISPR ...

...indicated, the antibiotic powder can be dissolved in dH 2 0. Addgene recommends making 1000X stock solutions...daughter cells. These daughter cells contain the same genetic information as the parental cell, and are...with the host cell. Like the bacterial chromosomal DNA, plasmid DNA is replicated upon cell division, and...chemistry enable isolation and purification of cellular components, such as DNA, but practically this ...and each daughter cell receives at least one copy of the plasmid. By the 1970s the combined discoveries...are thus termed clones of the original cell. The plasmid DNA is similarly referred to as cloned DNA, and... and ideally would replicate these plasmids upon cell division in addition to their own genomic DNA. Because...