Jove
Visualize
Contact Us
JoVE
x logofacebook logolinkedin logoyoutube logo
ABOUT JoVE
OverviewLeadershipBlogJoVE Help Center
AUTHORS
Publishing ProcessEditorial BoardScope & PoliciesPeer ReviewFAQSubmit
LIBRARIANS
TestimonialsSubscriptionsAccessResourcesLibrary Advisory BoardFAQ
RESEARCH
JoVE JournalMethods CollectionsJoVE Encyclopedia of ExperimentsArchive
EDUCATION
JoVE CoreJoVE BusinessJoVE Science EducationJoVE Lab ManualFaculty Resource CenterFaculty Site
Terms & Conditions of Use
Privacy Policy
Policies

Related Concept Videos

Scale-Up Processes01:14

Scale-Up Processes

The scale-up of microbial fermentation processes is essential in industrial biotechnology, allowing the transition from laboratory-scale experiments to commercial-scale production while aiming to maintain product yield and quality. This process requires meticulous adjustment of equipment design, process parameters, and contamination control strategies to accommodate increasing culture volumes.At the laboratory scale, cultures are typically maintained in 1 to 10-liter glass or autoclavable...
Upstream Processing01:27

Upstream Processing

Upstream processing represents a critical phase in biomanufacturing, wherein biological systems such as microorganisms, mammalian cells, or insect cells are cultivated to produce therapeutic proteins, vaccines, enzymes, or other biologically derived products. This phase encompasses all steps from the selection and genetic manipulation of the production organism to the cultivation of cells in bioreactors under tightly controlled environmental conditions.Host Selection and Genetic OptimizationThe...
Production of Antibiotics01:27

Production of Antibiotics

Penicillin, one of the earliest and most widely used antibiotics, is produced industrially by the filamentous fungus Penicillium chrysogenum. Large stirred-tank bioreactors ranging from tens to hundreds of thousands of liters maintain tightly controlled temperature, pH, and dissolved oxygen conditions to support fungal metabolism and maximize antibiotic yield. Penicillin is a secondary metabolite, synthesized primarily during the stationary growth phase, which requires a carefully managed...
Production of Pharmaceuticals01:30

Production of Pharmaceuticals

Industrial insulin production uses genetically engineered E. coli expressing a proinsulin gene controlled by a tryptophan promoter and containing a methionine linker for later cleavage. The cells also carry ampicillin resistance for selective growth. Seed cultures are stored at −80 °C and production begins by thawing a small amount to inoculate starter cultures, which are progressively scaled to a 50,000-L bioreactor. In the bioreactor, E. coli grow in nutrient-rich media under sterile, tightly...
Vaccine Production01:23

Vaccine Production

Vaccine production involves a sequence of upstream and downstream processes to generate a safe and effective immunological product. It begins with cultivating microorganisms, such as viruses or bacteria, to obtain antigenic material. For viral vaccines, mammalian host cells are grown in bioreactors and subsequently infected with the target virus. The virus replicates within the host cells, which are lysed to release viral particles. This lysate is then clarified through filtration or...

You might also read

Related Articles

Articles linked to this work by shared authors, journal, and citation graph.

Sort by
Same author

Emergence of fibrotic pericytes and their transcriptional regulation in pulmonary fibrosis.

Nature communications·2026
Same author

Biomimetic cellular sponges for neutralizing inflammatory cytokines in osteoarthritic joints.

Materials today. Bio·2026
Same author

Multifunctional ions/drugs co-delivering nanocomposite hydrogel orchestrates neuro-osteogenic microenvironment for boosting osteoporotic osseointegration.

Journal of nanobiotechnology·2026
Same author

FOXM1 inhibitor, RCM‑1, enhances venetoclax mediated apoptosis through downregulation of ATP2B4 in rhabdomyosarcoma.

International journal of oncology·2026
Same author

FOXF2 regulates pericyte-endothelial signaling required for vascular homeostasis after neonatal hyperoxic lung injury.

Nature communications·2026
Same author

Rebalancing NTRK2 isoforms promotes vascular regeneration in bronchopulmonary dysplasia.

Cell stem cell·2025

Related Experiment Video

Updated: Jul 20, 2026

Production of Double-stranded DNA Ministrings
06:12

Production of Double-stranded DNA Ministrings

Published on: February 29, 2016

10.4K

Protocol for minicircle production for gene therapy without subsequent cleanup steps.

Jonathan Do1, Gautam Verma1, Yasmin Maurice1

  • 1Phoenix Children's Research Institute, University of Arizona College of Medicine-Phoenix, Phoenix, AZ, USA.

STAR Protocols
|July 25, 2025
PubMed
Summary

This study introduces a streamlined protocol to significantly boost minicircle DNA production, achieving up to a 10-fold increase by removing intermediate cleanup steps for enhanced yields.

Keywords:
Biotechnology and bioengineeringGene ExpressionMicrobiology

More Related Videos

Characterization of Functionally Associated miRNAs in Glioblastoma and their Engineering into Artificial Clusters for Gene Therapy
09:40

Characterization of Functionally Associated miRNAs in Glioblastoma and their Engineering into Artificial Clusters for Gene Therapy

Published on: October 4, 2019

5.7K
CIRCLE-Seq for Interrogation of Off-Target Gene Editing
08:23

CIRCLE-Seq for Interrogation of Off-Target Gene Editing

Published on: November 1, 2024

861

Related Experiment Videos

Last Updated: Jul 20, 2026

Production of Double-stranded DNA Ministrings
06:12

Production of Double-stranded DNA Ministrings

Published on: February 29, 2016

10.4K
Characterization of Functionally Associated miRNAs in Glioblastoma and their Engineering into Artificial Clusters for Gene Therapy
09:40

Characterization of Functionally Associated miRNAs in Glioblastoma and their Engineering into Artificial Clusters for Gene Therapy

Published on: October 4, 2019

5.7K
CIRCLE-Seq for Interrogation of Off-Target Gene Editing
08:23

CIRCLE-Seq for Interrogation of Off-Target Gene Editing

Published on: November 1, 2024

861

Area of Science:

  • Molecular Biology
  • Biotechnology
  • Genetic Engineering

Background:

  • Minicircle DNA production is crucial for various biotechnological applications, including gene therapy and vaccine development.
  • Existing methods for minicircle DNA synthesis often suffer from low yields due to extensive purification procedures.
  • Optimizing minicircle production is essential for cost-effective and scalable downstream applications.

Purpose of the Study:

  • To develop an improved protocol for high-yield minicircle DNA production.
  • To eliminate post-induction cleanup steps to streamline the process and increase efficiency.
  • To demonstrate a significant increase in minicircle yield compared to current techniques.

Main Methods:

  • The protocol involves optimized bacterial culture and same-day arabinose induction.
  • DNA harvesting is performed without intermediate purification steps.
  • The method is applicable to minicircles derived from parental plasmids with specific attB, attP, and multiple Isce-I endonuclease sites in ZYCY10PS3T2 bacteria.

Main Results:

  • The presented protocol increases minicircle DNA yield by up to 10-fold.
  • Elimination of post-induction cleanup steps significantly enhances production efficiency.
  • The method provides a robust and scalable approach for minicircle generation.

Conclusions:

  • This optimized protocol offers a substantial improvement in minicircle DNA yield.
  • The simplified procedure reduces processing time and resource requirements.
  • The findings facilitate broader application of minicircle DNA technology in research and therapeutics.