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October 3, 2019Journal of Visualized ExperimentsOpen Access

Lentiviral CRISPR/Cas9 editing achieves ~90% myeloid cell transduction at four weeks in mice.

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Why the study?

Manipulating genes in hematopoietic stem cells using conventional transgenesis approaches can be time-consuming, expensive, and challenging.

Population

Lethally-irradiated C57BL/6 mice transplanted with Cas9-expressing lineage-negative bone marrow cells

Comparison

Targeting gRNA and RFP vs non-targeting gRNA

Design

Preclinical experimental protocol

Follow-up

4 weeks after transplantation

Key result

Lentiviral CRISPR/Cas9-mediated genome editing achieved ~90% transduction of myeloid cells and ~70% of lymphoid cells at 4 weeks after transplantation in mice.

Authors

SSSoichi SanoYWYing WangMEMegan A. Evans

Discussion

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Overview

Enables efficient HSC gene editing in mice; leaves open translation to human applications.

Structured PICO

P
Population
Cas9-expressing lineage-negative bone marrow cells transplanted into lethally-irradiated C57BL/6 mice
I
Intervention
Lentiviral transduction with a guide RNA (gRNA) targeting specific genes and a red fluorescence reporter gene (RFP)
C
Comparator
Mice transplanted with lentivirus expressing non-targeting gRNA
O
Outcome
Engraftment of transduced hematopoietic stem cells evaluated by flow cytometric analysis of RFP-positive leukocytes of peripheral bloodsurrogate

This protocol provides an efficient and economical method for high-throughput evaluation of hematopoiesis-regulatory genes in mouse disease models.

Cite This Study

Sano et al. (2019) studied Hematopoietic cells in disease models. Lentiviral CRISPR/Cas9-mediated genome editing vs. Lentivirus expressing non-targeting gRNA was evaluated on Transduction efficiency of myeloid and lymphoid cells. Lentiviral CRISPR/Cas9-mediated genome editing achieved ~90% transduction of myeloid cells and ~70% of lymphoid cells at 4 weeks after transplantation in mice.

synapsesocial.com/papers/6aa93debd0c7c778ac8cc37dhttps://doi.org/10.3791/59977
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Also Consider

Synapse has enriched 5 closely related papers on similar clinical questions. Consider them for comparative context:

  1. 1In situ gene editing of hematopoietic stem cells via AAV-delivered CRISPR guide RNAs2025 · 3 citations
  2. 2Gene-edited hematopoietic stem cells for leukemia and lymphoma treatment: a systematic review of preclinical and translational evidence2025 · 4 citations
  3. 3Transgenic expression of enhanced AsCas12a in mice enables In Vivo multiplex genetic editing and development of diverse novel models of myeloid neoplasms2025
  4. 4CRISPR-Cas9 Mediated Gene Editing in Inherited Hematologic Disorders2025
  5. 5Engraftment Outcome of CRISPR/Cas9-Edited Hematopoietic Stem Cells for Genetic Diseases: A Systematic Review and Meta-Analysis of Preclinical Evidence2026