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

CRISPR/Cas9 Genome Editing01:28

CRISPR/Cas9 Genome Editing

1.4K
The CRISPR-Cas system serves as a bacterial defense mechanism against invading genetic elements such as viruses and plasmids, forming the foundation for its adaptation as a powerful genome-editing tool. Originally discovered in prokaryotes, this system has been repurposed to revolutionize genetic engineering across a wide range of organisms, including plants, animals, and humans. The core component, Cas9, is an endonuclease derived from Streptococcus pyogenes, capable of introducing...
1.4K
CRISPR01:59

CRISPR

57.2K
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...
57.2K

You might also read

Related Articles

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

Sort by
Same author

SpatioTemporal Omics Consortium: a global effort for biological discovery across species, space and time.

Nature methods·2026
Same author

EasySCP unveils extensive liver zonation at single-cell proteomics resolution.

Nature communications·2026
Same author

Modeling and correction of SCID-X1 using CRISPR-Cas9 homology-directed repair in human HSPCs.

Molecular therapy. Nucleic acids·2026
Same author

Clinical development of tacrolimus-resistant regulatory T cells to enable simultaneous immunosuppression and immune regulation.

Molecular therapy. Advances·2026
Same author

TMPRSS2-ERG confers resistance of prostate cancer to antiandrogens.

EMBO molecular medicine·2026
Same author

Development of a Fully Non-Viral 1XX-enhanced BCMA CAR-T Cell Therapy for Multiple Myeloma.

bioRxiv : the preprint server for biology·2026

Related Experiment Video

Updated: Dec 20, 2025

Genome Editing and Directed Differentiation of hPSCs for Interrogating Lineage Determinants in Human Pancreatic Development
09:37

Genome Editing and Directed Differentiation of hPSCs for Interrogating Lineage Determinants in Human Pancreatic Development

Published on: March 5, 2017

13.4K

Increasing CRISPR Efficiency and Measuring Its Specificity in HSPCs Using a Clinically Relevant System.

Jenny Shapiro1, Ortal Iancu1, Ashley M Jacobi2

  • 1Institute of Nanotechnology and Advanced Materials, The Mina and Everard Goodman Faculty of Life Sciences, Bar-Ilan University, Ramat-Gan 5290002, Israel.

Molecular Therapy. Methods & Clinical Development
|June 2, 2020
PubMed
Summary
This summary is machine-generated.

Optimizing CRISPR-Cas9 genome editing in hematopoietic stem cells (HSPCs) enhances therapeutic potential. This study refined ex vivo editing techniques and established methods for identifying off-target effects in CD34+ HSPCs.

Keywords:
CD34+ hematopoietic stem and progenitor cellsCRISPR-Cas9chemically modified guide RNAsgenome editingoff-target sites

More Related Videos

Author Spotlight: Development of Simplified CRISPR-Based Tests for Rapid Detection of Infectious Diseases
10:16

Author Spotlight: Development of Simplified CRISPR-Based Tests for Rapid Detection of Infectious Diseases

Published on: August 16, 2024

1.9K
Genome-Wide CRISPR Screen for Unveiling Radiosensitive and Radioresistant Genes
08:32

Genome-Wide CRISPR Screen for Unveiling Radiosensitive and Radioresistant Genes

Published on: May 23, 2025

936

Related Experiment Videos

Last Updated: Dec 20, 2025

Genome Editing and Directed Differentiation of hPSCs for Interrogating Lineage Determinants in Human Pancreatic Development
09:37

Genome Editing and Directed Differentiation of hPSCs for Interrogating Lineage Determinants in Human Pancreatic Development

Published on: March 5, 2017

13.4K
Author Spotlight: Development of Simplified CRISPR-Based Tests for Rapid Detection of Infectious Diseases
10:16

Author Spotlight: Development of Simplified CRISPR-Based Tests for Rapid Detection of Infectious Diseases

Published on: August 16, 2024

1.9K
Genome-Wide CRISPR Screen for Unveiling Radiosensitive and Radioresistant Genes
08:32

Genome-Wide CRISPR Screen for Unveiling Radiosensitive and Radioresistant Genes

Published on: May 23, 2025

936

Area of Science:

  • Molecular Biology
  • Gene Editing Technologies
  • Hematopoietic Stem Cell Biology

Background:

  • Hematopoietic stem and progenitor cells (HSPCs) are crucial for treating blood disorders.
  • CRISPR-Cas9 genome editing offers therapeutic promise for HSPCs but requires optimization.
  • Precise control and off-target assessment are vital for clinical translation.

Purpose of the Study:

  • To optimize ex vivo CRISPR-Cas9 genome editing efficiency in human CD34+ HSPCs.
  • To develop a robust workflow for identifying and quantifying off-target mutations.
  • To evaluate different guide RNA formats and delivery methods for therapeutic applications.

Main Methods:

  • Utilized CRISPR-Cas9 ribonucleoprotein (RNP) complexes with modified synthetic guide RNAs (gRNAs) in CD34+ HSPCs.
  • Investigated the impact of Alt-R electroporation enhancer (EE) and excess gRNA on editing efficiency.
  • Employed Genome-Wide, Unbiased Identification of Double-Strand Breaks Enabled by Sequencing (GUIDE-seq) for off-target analysis.
  • Quantified editing using rhAmpSeq technology.

Main Results:

  • Addition of an electroporation enhancer (EE) significantly increased editing efficiency in CD34+ HSPCs.
  • Using excess guide RNA (gRNA) over Cas9 protein also improved editing, offering a DNA-free alternative.
  • Single-guide RNA (sgRNA) demonstrated potential advantages over 2-part gRNA in a locus-specific manner.
  • Established a comprehensive framework for unbiased off-target site identification and quantification.

Conclusions:

  • Optimized CRISPR-Cas9 editing protocols enhance efficiency in CD34+ HSPCs.
  • Developed reliable methods for assessing genome editing safety and specificity.
  • Findings support the clinical translation of genome editing for hematopoietic stem cell therapies.