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Related Concept Videos

DNA Damage can Stall the Cell Cycle02:36

DNA Damage can Stall the Cell Cycle

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In response to DNA damage, cells can pause the cell cycle to assess and repair the breaks. However, the cell must check the DNA at certain critical stages during the cell cycle. If the cell cycle pauses before DNA replication, the cells will contain twice the amount of DNA. On the other hand, if cells arrest after DNA replication but before mitosis, they will contain four times the normal amount of DNA. With a host of specialized proteins at their disposal,cells must use the right protein at...
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DNA Damage Can Stall the Cell Cycle02:36

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In response to DNA damage, cells can pause the cell cycle to assess and repair the breaks. However, the cell must check the DNA at certain critical stages during the cell cycle. If the cell cycle pauses before DNA replication, the cells will contain twice the amount of DNA. On the other hand, if cells arrest after DNA replication but before mitosis, they will contain four times the normal amount of DNA. With a host of specialized proteins at their disposal,cells must use the right protein at...
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Negative Regulator Molecules01:23

Negative Regulator Molecules

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Positive regulators allow a cell to advance through cell cycle checkpoints. Negative regulators have an equally important role as they terminate a cell’s progression through the cell cycle—or pause it—until the cell meets specific criteria.
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The Cell Cycle Control System01:28

The Cell Cycle Control System

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The cell cycle regulation directs how a cell proceeds from one phase to the next and begins mitosis. The cell cycle control system includes intracellular regulatory molecules and external triggers. They provide "stop" or "advance" signals and operate at specific cell cycle stages termed checkpoints to ensure that a particular process is completed before the cell advances to the next phase.
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Anaphase Promoting Complex00:50

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The stepwise destruction of specific proteins is necessary for the progression and completion of the cell cycle. Such proteins are ubiquitinated by ubiquitin ligases and then subsequently destroyed by the proteasome. The SCF (Skp1/Cullin/F-box) and the anaphase-promoting complex (APC) are two important ubiquitin ligases involved in cell cycle progression. While SCF is active throughout the cell cycle, APC gets activated during metaphase to anaphase transition. Cdc20 or Cdh1 binds to APC and...
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Inhibition of Cdk Activity02:34

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The orderly progression of the cell cycle depends on the activation of Cdk protein by binding to its cyclin partner. However, the cell cycle must be restricted when undergoing abnormal changes. Most cancers correlate to the deregulated cell cycle, and since Cdks are a central component of the cell cycle, Cdk inhibitors are extensively studied to develop anticancer agents. For instance, cyclin D associates with several Cdks, such as Cdk 4/6, to form an active complex. The cyclin D-Cdk4/6 complex...
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Production, Purification, and Quality Control for Adeno-associated Virus-based Vectors
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AAV Assembled Capsids Are Produced in Cells Blocked From Cell Cycle Progression.

Alaka Mullick1, Audrey Morasse1, Melanie Leclerc1

  • 1Human Health Therapeutics, National Research Council of Canada, Montreal, Quebec, Canada.

Biotechnology and Bioengineering
|November 19, 2025
PubMed
Summary

Gene therapy production of adeno-associated virus (AAV) is inefficient. We found that cells successfully transfected with AAV DNA can be blocked in their cell cycle, impacting capsid production.

Keywords:
AAVE1ARNA‐seqadeno‐associated viruscell cycle

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A Quantitative Dot Blot Assay for AAV Titration and Its Use for Functional Assessment of the Adeno-associated Virus Assembly-activating Proteins
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Process Development for the Production and Purification of Adeno-Associated Virus AAV2 Vector using Baculovirus-Insect Cell Culture System
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Process Development for the Production and Purification of Adeno-Associated Virus AAV2 Vector using Baculovirus-Insect Cell Culture System

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Area of Science:

  • Biotechnology
  • Molecular Biology
  • Gene Therapy

Background:

  • Adeno-associated virus (AAV) is a key vector for gene therapy.
  • Current AAV manufacturing yields are low, leading to high costs and limited patient access.
  • A significant portion of transfected cells fail to produce AAV capsids.

Purpose of the Study:

  • To investigate why some cells transfected with AAV DNA do not produce assembled capsids.
  • To characterize the differences between AAV-producing and non-producing cell populations.
  • To identify molecular targets for improving AAV production efficiency.

Main Methods:

  • Cellular characterization of transfected HEK293 cells.
  • Cell cycle analysis of producer and non-producer populations.
  • RNA sequencing (RNA-seq) to compare gene expression profiles.

Main Results:

  • AAV-producing cells exhibited a distinct cell cycle arrest compared to non-producers.
  • RNA-seq revealed significant differences in molecular pathways between the two cell populations.
  • These molecular differences provide insights into factors limiting AAV capsid assembly.

Conclusions:

  • Cell cycle progression is a critical factor influencing AAV production in transfected cells.
  • Understanding the molecular basis of this block can guide strategies to enhance AAV manufacturing yields.
  • Targeting cell cycle regulation may overcome limitations in current gene therapy vector production.