...expression from the other episomal plasmids. When used on PBMCs, the pCXLE episomal vector system increases...non-integrating RNA virus Episomal vectors- extrachromosomally maintained DNA plasmids that express reprogramming...Reprogramming efficiency of PBMCs with episomal vectors Episomal vectors are based on the Epstein-Barr ... in human cells [8]. The EBNA-1 protein recognizes OriP and induces amplification of the episomes concurrently... pCXLE episomal vector system generated by the group is an optimized mixture of three episomal plasmids...available at Addgene. Table 1: List of pCXLE episomal plasmids, their Addgene ID numbers and the reprogramming...reprogramming factors they encode Episomal Plasmid Addgene ID# Encodes pCXLE-hSK 27078 SOX2 and KLF4 ...

...into cells as a donor plasmid containing the transposon, along with a helper plasmid that expresses the ...reprogramming factors. Non-replicating Plasmids: Generating iPSCs with plasmid-based expression requires serial... generating iPSCs by plasmid transfection where 2 of 11 clones tested had plasmid integration. Another...the dose of plasmid the cells receive over the whole reprogramming period and the plasmid will get diluted...larger plasmids have lower rates of transfection. Minicircles: Minicircles are like mini-plasmids. They...Epstein-Barr nuclear antigen-1 (EBNA1) based episomes: These plasmids carry the origin of replication (oriP)...reflection of the large size of the plasmid or silencing of the plasmid by DNA methylation. Another downside...

...genome. AAV instead relies mainly on episomal expression. Episomes are circular pieces of DNA that reside... The most obvious workaround is to use a multi-plasmid system, where you have one AAV vector dedicated... R. J. (2022). AAV vectors: The Rubik’s cube of human gene therapy. Molecular Therapy, 30(12), 3515–3541...& Kampmann, M. (2023). CRISPR screening by AAV episome-sequencing (CrAAVe-seq) is a highly scalable cell... virus as a delivery vector for gene therapy of human diseases. Signal Transduction and Targeted Therapy...

...addition of a helper virus or plasmid allows it to replicate episomally, essentially remaining an extrachromosomal...cancer-connection has been identified in humans to date. Over fifty serotypes of human-infecting adenovirus have been...their required components are very similar. While plasmids containing virus-specific elements such as the... single-stranded DNA parvovirus. AAV can infect humans and some other mammals but is not currently known...Altogether, AAV has many excellent features for human gene therapy and is currently being used in clinical...DNA genome which is non-enveloped. It can infect humans, many mammals, and even the avian population. Some...-associated Virus (AAV) Guide Addgene’s viral plasmids and resources Addgene’s viral services: AAV ...

...Gonçalves and Ventura labs’ plasmids, including protocols, check out the plasmids at Addgene: pAdSh.PGK.Cas9...looking for a broader range of CRISPRs plasmid tools, find more plasmids, CRISPR technology guides, FAQs, and...donor DNA leads to a vast majority of AdV-modified human cells being subjected to scarless homology-directed...description of their experimental protocol. The three plasmids which have been deposited to Addgene are: pAdSh.PGK.Cas9...Gonçalves says that advantages of AdVs include their episomal nature and very efficient introduction of DNA ...induces targeted mutagenesis in a diverse array of human cells. Sci Rep 4: . https://doi.org/10.1038/srep05105...Liver in Spite of Cas9-Specific Immune Responses. Human Gene Therapy 26:432–442 . https://doi.org/10.1089...

... an organism. These features make AAV ideal for human therapies and in vivo laboratory experiments ranging...host genome and therefore the transduced gene is episomal and does not divide and replicate in the same ...of both polarities are rescued from the vector plasmid and individually packaged into the AAV capsids....medium during adeno-associated virus manufacturing." Human gene therapy 21.10 (2010): 1251-1257. PubMed PMID...

...becoming available. At the same time advances in human genetics led to the identification of several disease-causing...visual impairment that faithfully recapitulated the human condition. Luck brought all these key elements together...therapy. It is not known to cause any disease in humans, and its low immunogenicity is critical when systemic...expression in non-dividing cells because they remain in episomal form in the cell’s nucleus. But since nothing...in mammals Resources at Addgene.org Find AAV plasmids Find ready-to-use AAV viral preps Browse all viral...

... [6], plasmid electroporation (episomal) [7, 8] and mRNA transfection [9]. Many of the plasmids used for...easily isolated from donors. Apart from fibroblasts, human keratinocytes from hair pluck, peripheral blood ...types have been successfully differentiated from human iPSCs. Below is a list of some of these cell types...Neurons [32, 33, 34] You can find a variety of plasmids for differentiation here. Transdifferentiation... Induction of pluripotent stem cells from adult human fibroblasts by defined factors. Cell, 2007. 131(...Induced pluripotent stem cell lines derived from human somatic cells. Science, 2007. 318(5858): p. 1917... et al., Efficient generation of transgene-free human induced pluripotent stem cells (iPSCs) by temperature-sensitive...