Episomal Vectors
Episomal Vectors - Related Videos
09:45
Generation of Integration-free Induced Pluripotent Stem Cells from Human Peripheral Blood Mononuclear Cells Using Episomal Vectors
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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
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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.
10:52
Generation of Induced Pluripotent Stem Cells from Frozen Buffy Coats using Non-integrating Episomal Plasmids
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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.
09:20
Vectors in Multiple Directions
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2023
Source: Nicholas Timmons, Asantha Cooray, PhD, Department of Physics & Astronomy, School of Physical Sciences, University of California, Irvine, CA This experiment demonstrates how vectors add and subtract in multiple directions. The goal will be to analytically calculate the addition or subtraction of multiple vectors and then to experimentally confirm the calculations. A vector is an object with both magnitude and direction. The magnitude of a vector is simply denoted as the length,...
10:42
A Protocol for the Production of Integrase-deficient Lentiviral Vectors for CRISPR/Cas9-mediated Gene Knockout in Dividing Cells
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Cited by 21 •
2017
We describe the production strategy of integrase-deficient lentiviral vectors (IDLVs) as vehicles for delivering CRISPR/Cas9 to cells. With an ability to mediate quick and robust gene editing in cells, IDLVs present a safer and equally effective vector platform for gene delivery compared to integrase-competent vectors.
FAQs
Regulatory elements determine when and how the genetic cargo is transcribed after an episomal vector enters a cell. By directing transcription rather than relying on chromosomal insertion, they help researchers control cargo expression and support potentially reversible experiments. This design is especially relevant when bioengineers need protein production or cellular changes without making the alteration permanent.
Persistence depends on more than cargo expression. In some systems, viral origins provide a basis for episome replication, while maintenance proteins support retention of the episomal DNA. If those mechanisms are absent or insufficient, cell division can dilute the DNA among daughter cells, reducing the duration of expression. Replication and maintenance therefore influence whether activity remains brief or continues.
The key distinction is where the genetic material remains. Episomal designs keep cargo outside host chromosomes, whereas integrating approaches place it into the genome. This difference makes episomal delivery attractive when permanent genomic alteration is undesirable, although expression may decline as cells divide if replication and maintenance mechanisms do not preserve the episome.
Expression outcomes reflect several linked steps: vector entry, activity of regulatory elements, and, in suitable systems, episome replication and maintenance. Entry and transcription enable initial production, whereas replication and maintenance support persistence across cell divisions. When the latter processes are unavailable, dilution can shorten the period of detectable cargo expression, even if transcription begins successfully.
A basic bioengineering workflow begins with delivery of the episomal DNA into cells, followed by transcription of the chosen cargo from the vector’s regulatory elements. Researchers then consider whether the system includes viral origins and maintenance proteins, because these features affect persistence. The resulting expression may be used while avoiding a planned permanent chromosomal change.
Episomal systems support several applications named in the overview: transient protein production, cellular reprogramming, genome engineering, and cell therapy research. Their value differs by goal. Temporary expression can provide a limited period of cargo activity, while nonintegration is important in studies where researchers want to reduce permanent genomic alteration. These features support controlled experimental designs.
A decline in expression after cell division can indicate dilution of episomal DNA rather than failure of transcription at the outset. Researchers can distinguish initial cargo activity from longer-term persistence by considering whether the system contains replication and maintenance features. This interpretation helps explain why a vector may support transient output but not sustained expression in dividing cells.