ArticleJournal of visualized experiments : JoVE2019
Lentiviral CRISPR/Cas9-Mediated Genome Editing for the Study of Hematopoietic Cells in Disease Models.
Article in Journal of visualized experiments : JoVE, 2019. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Cited by 14 papers.
What it found
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The abstract states no effect estimate the extractor could read, or names no intervention and outcome on the map, so this paper lights no cell and moves no belief. It is still indexed, cited and linked below.
The trial behind it
Trials whose registry record cites this paper, or whose number appears in the abstract. A trial that started after this paper was published is citing it as background, not reporting it.
Neither the registry nor the abstract names a trial number. If this is a trial report, that itself is worth knowing.
Who cites it
14 citing papers in PubMed, 14 citations in OpenAlex.
- Disruption of theNature cardiovascular research · 2024Article
- Development of a Mouse Model of Hematopoietic Loss of Y Chromosome.Bio-protocol · 2023Article
- Circulating macrophages as the mechanistic link between mosaic loss of Y-chromosome and cardiac disease.Cell & bioscience · 2023Article
- Clonal Hematopoiesis: Connecting Aging and Inflammation in Atherosclerosis.Current atherosclerosis reports · 2023Review
- Clonal hematopoiesis, somatic mosaicism, and age-associated disease.Physiological reviews · 2023Review
- Advances in CRISPR Delivery Methods: Perspectives and Challenges.The CRISPR journal · 2022Review
- Hematopoietic loss of Y chromosome leads to cardiac fibrosis and heart failure mortality.Science (New York, N.Y.) · 2022Article
- Murine models of clonal haematopoiesis to assess mechanisms of cardiovascular disease.Cardiovascular research · 2022Review
- Importance of clonal hematopoiesis in heart failure.Trends in cardiovascular medicine · 2022Review
- Evidence for reciprocal network interactions between injured hearts and cancer.Frontiers in cardiovascular medicine · 2022Review
- The Cancer Therapy-Related Clonal Hematopoiesis Driver GeneCirculation research · 2021Article
- Employing the CRISPR-Cas System for Clonal Hematopoiesis Research.International journal of physical medicine & rehabilitation · 2021Article
- The role of clonal haematopoiesis in cardiovascular diseases: epidemiology and experimental studies.Journal of internal medicine · 2020Review
- Article
Corrections and comments
PubMed lists nothing against this paper. Absence here is not a guarantee, only a check that was made.
Authors and funding
9 authors at 1 institution in 1 country.
Funding
Abstract
Manipulating genes in hematopoietic stem cells using conventional transgenesis approaches can be time-consuming, expensive, and challenging. Benefiting from advances in genome editing technology and lentivirus-mediated transgene delivery systems, an efficient and economical method is described here that establishes mice in which genes are manipulated specifically in hematopoietic stem cells. Lentiviruses are used to transduce Cas9-expressing lineage-negative bone marrow cells with a guide RNA (gRNA) targeting specific genes and a red fluorescence reporter gene (RFP), then these cells are transplanted into lethally-irradiated C57BL/6 mice. Mice transplanted with lentivirus expressing non-targeting gRNA are used as controls. Engraftment of transduced hematopoietic stem cells are evaluated by flow cytometric analysis of RFP-positive leukocytes of peripheral blood. Using this method, ~90% transduction of myeloid cells and ~70% of lymphoid cells at 4 weeks after transplantation can be achieved. Genomic DNA is isolated from RFP-positive blood cells, and portions of the targeted site DNA are amplified by PCR to validate the genome editing. This protocol provides a high-throughput evaluation of hematopoiesis-regulatory genes and can be extended to a variety of mouse disease models with hematopoietic cell involvement.
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Registered trials
Read under generation 80e0d062 · epoch 390. Bibliography from PubMed, PubMed Central and OpenAlex; grants from NIH RePORTER; trial links from ClinicalTrials.gov; estimates, votes and beliefs from the OpenQuestion graph.