Sv40 Episome
Sv40 Episome - Related Videos
05:22
Electrophoretic Analysis of Replication Through Structure-Prone DNA Repeats Within the SV40-Based Human Episome
0 Views •
2024
Here, we outline the procedure for analyzing replication progression through pathogenic, structure-prone repeats using 2-dimensional gel electrophoresis.
10:52
Generation of Induced Pluripotent Stem Cells from Frozen Buffy Coats using Non-integrating Episomal Plasmids
0 Views •
Cited by 26 •
2015
Induced pluripotent stem cells (iPSCs) represent a source of patient-specific tissues for clinical applications and basic research. Here, we present a detailed protocol to reprogram human peripheral blood mononuclear cells (PBMNCs) obtained from frozen buffy coats into viral-free iPSCs using non-integrating episomal plasmids.
09:45
Generation of Integration-free Induced Pluripotent Stem Cells from Human Peripheral Blood Mononuclear Cells Using Episomal Vectors
0 Views •
Cited by 17 •
2017
This protocol describes a detailed method for efficient generation of integration-free iPSCs from human adult peripheral blood cells. With the use of four oriP/EBNA-based episomal vectors to express the reprogramming factors, KLF4, MYC, BCL-XL, or OCT4 and SOX2, thousands of iPSC colonies can be obtained from 1 mL of peripheral blood.
09:56
Epigenetic Engineering of K562 Cells: Dual-Vector Episomal Strategy for Stable Targeted DNA Methylation using dCas9-DNMT3A and -HDAC1 Fusion Proteins
0 Views •
2025
This protocol describes a non-integrative, episomal CRISPR/dCas9-based system for targeted epigenetic editing in K562 cells, combining dCas9-DNMT3A and dCas9-HDAC1 effectors with specific sgRNAs to induce locus-specific DNA methylation with precision and reduced off-target effects.
08:14
Efficient iPS Cell Generation from Blood Using Episomes and HDAC Inhibitors
0 Views •
Cited by 18 •
2014
Here we describe a protocol for generating human induced pluripotent stem cells from peripheral blood using an episome based reprogramming strategy and histone deacetylase inhibitors.
FAQs
Replication begins when SV40 large T antigen recognizes and binds the viral replication origin carried by the episome. This interaction promotes DNA unwinding, creating access for replication enzymes supplied by the mammalian host cell. The system therefore combines a virus-derived initiation factor with cellular replication machinery, making it useful for examining how these activities support extrachromosomal DNA maintenance.
The SV40 replication origin provides the specific DNA site recognized by large T antigen. Without this origin, the viral initiation mechanism described for the episome cannot be engaged. Its presence allows researchers to connect origin recognition, DNA unwinding, and host-enzyme activity with the persistence of an extrachromosomal molecule, helping clarify how replication control influences episomal behavior.
Persistence is influenced by what happens to episomal DNA as host cells divide. Even when the molecule remains outside the chromosomes, its abundance can change through dilution during cell division. Monitoring persistence and dilution helps researchers distinguish continued maintenance from gradual loss and evaluate how effectively an episomal system remains available for studies of gene expression or DNA organization.
An SV40 episome can support gene expression without immediate insertion into the host chromosomes, whereas chromosomal integration places genetic material within the genome. This distinction lets investigators study nonintegrating genetic-vector behavior and compare transient or persistent extrachromosomal activity with the consequences of genomic insertion, including differences in persistence, dilution, and effects on host-cell biology.
These systems support investigations of DNA replication, plasmid maintenance, chromatin organization, and regulation of gene expression. Because the DNA remains separate from the chromosomes, researchers can examine how an extrachromosomal molecule is replicated and maintained while also assessing its influence on cellular regulatory processes. The approach connects molecular replication events with broader genetic and chromatin-related outcomes.
Evaluation should include whether the episomal DNA persists, how its abundance changes through cell division, and whether it affects host-cell biology. Researchers can also examine gene-expression behavior, plasmid maintenance, chromatin organization, and replication activity. Together, these measurements show whether the system provides a useful nonintegrating platform and reveal tradeoffs between persistence and progressive dilution.