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Related Concept Videos

CRISPR01:59

CRISPR

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Genome editing technologies allow scientists to modify an organism’s DNA via the addition, removal, or rearrangement of genetic material at specific genomic locations. These types of techniques could potentially be used to cure genetic disorders such as hemophilia and sickle cell anemia. One popular and widely used DNA-editing research tool that could lead to safe and effective cures for genetic disorders is the CRISPR-Cas9 system. CRISPR-Cas9 stands for Clustered Regularly Interspaced...
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Genome Editing and Directed Differentiation of hPSCs for Interrogating Lineage Determinants in Human Pancreatic Development
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Toward the Development of a Tissue Engineered Gradient Utilizing CRISPR-Guided Gene Modulation.

Jacob D Weston1, Brooke Austin1, Hunter Levis1

  • 1Department of Biomedical Engineering, University of Utah, Salt Lake City, Utah, USA.

Tissue Engineering. Part A
|February 7, 2024
PubMed
Summary
This summary is machine-generated.

CRISPR gene modulation directs stem cell differentiation for tissue engineering. This approach successfully created cellular and tissue gradients without growth factors, mimicking native tissue structures.

Keywords:
ASCsCRISPRaCRISPRienthesisnogginosteogenesis

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Area of Science:

  • Biomaterials Science
  • Stem Cell Biology
  • Gene Editing Technologies

Background:

  • Biological tissues exhibit complex gradients in cellular, compositional, and mechanical properties, particularly at transition zones.
  • Engineering these native-like gradients in tissues remains challenging due to the intricate nature of biological systems.
  • Current tissue engineering strategies often rely on stem cells, which require precise environmental cues for differentiation, posing control difficulties in vitro and in vivo.

Purpose of the Study:

  • To demonstrate the effectiveness of clustered regularly-interspaced short palindromic repeats (CRISPR)-guided gene modulation in directing stem cell differentiation for tissue engineering.
  • To engineer cellular and tissue gradients using CRISPR-modulated adipose-derived stem cells (ASCs) without exogenous growth factors.
  • To create a proof-of-concept gradient mimicking the fibrocartilage-to-mineralized-fibrocartilage transition found in the enthesis.

Main Methods:

  • Screening of CRISPR-interference (CRISPRi) constructs targeting osteogenic inhibitor promoters in ASCs.
  • Utilizing CRISPRi to modulate Noggin expression for regulating osteogenic differentiation and mineral deposition.
  • Combining CRISPR-activation multiplex-engineered chondrogenic ASCs with a type I collagen construct for controlled cell deposition and gradient formation.

Main Results:

  • CRISPRi targeting of Noggin successfully regulated ASC osteogenic differentiation and mineral deposition without exogenous growth factors.
  • Demonstrated controlled deposition of engineered ASCs to form a gradient on an anisotropic collagen scaffold.
  • Successfully created a cell and tissue gradient analogous to native fibrocartilage-to-mineralized-fibrocartilage transitions.

Conclusions:

  • CRISPR-engineered ASCs offer a promising tool for creating complex tissue gradients.
  • This gene modulation strategy enables the development of engineered tissues with native-like gradients without relying on external growth factors.
  • The study highlights the potential of CRISPR technology in advancing tissue engineering applications for regenerative medicine.