Article Sidebar

PDF (Tiếng Việt)
Published: 2023-02-28
Abstract Views: 505
Views PDF (Tiếng Việt): 535
DOI: https://doi.org/10.52852/tcncyh.v163i2.1387
Issue
Vol. 163 No. 2 (2023)
Section
Các bài báo
How to Cite
Hà, V. T. ., Phượng, Đoàn T. K., Anh, L. T. L. ., Long, N. V., Anh, B. H., Bắc, Đào N. ., Trang, T. T. H., Trung, N. V., & Phấn, T. Đức. (2023). 4. A study on gene editing by crispr/cas9 system in hematopoietic stem cell. Journal of Medical Research, 163(2), 30-36. https://doi.org/10.52852/tcncyh.v163i2.1387

4. A study on gene editing by crispr/cas9 system in hematopoietic stem cell

Vu Thi Ha, Doan Thi Kim Phuong, Luong Thi Lan Anh, Nguyen Van Long, Bach Huy Anh, Dao Ngoc Bac, Tran Thi Huyen Trang, Nguyen Viet Trung, Tran Duc Phan

Main Article Content

Abstract

Hematopoietic stem cells are cells with many potential clinical applications for the treatment of blood diseases as well as cancers and autoimmune diseases treatment in general. Collecting, editing genes in stem cells, culturing, and proliferating stem cells as a premise for clinical trials are still a challenge of medicine. The Clustered Regularly Interspaced Short Palindromic Repeats and CRISPR-associated protein system is a highly efficient and promising gene editing tool for clinical application to control genes of interest. Stem cells are collected based on surface marker CD34+CD38- by flow cytometry system, targeted gene editing, mutated clone were cultured and proliferated through the aforementioned system. Result showed that ratio of hematopoietic stem cell in the cord blood cell were 0.3%. The efficiency of Spred1 editing by CRISPR/Cas9 systerm occured at the DNA and protein level. The research has successfully collected stem cells, stem cells after being edited with target genes proliferate, differentiate into independent cell lines and become a premise for further studies.

Article Details

Keywords

Hematopoietic stem cell, CRISPR/Cas9, gene therapy, human cord blood cell

References

1. Yao CL, Hwang SM. Ex vivo expansion of hematopoietic stem cells from human cord blood in serum-free conditions. Methods Mol Biol Clifton NJ. 2007; 407: 165-175. doi: 10.1007/978-1-59745-536-7_13.
2. Gundry MC, Brunetti L, Lin A, Mayle AE, Kitano A, Wagner D, Hsu JI, Hoegenauer KA, Rooney CM, Goodell MA, Nakada D. Highly Efficient Genome Editing of Murine and Human Hematopoietic Progenitor Cells by CRISPR/Cas9. Cell Rep. 2016 Oct 25; 17(5): 1453-1461. doi: 10.1016/j.celrep.2016.09.092. PMID: 27783956; PMCID: PMC5087995.
3. Ferrari Samuele, Vavassori Valentina, et al. Gene Editing of Hematopoietic Stem Cells: Hopes and Hurdles Toward Clinical Translation. Frontiers in Genome Editing. 2021; (3):10.3389/fgeed.2021.618378. 2673-3439.
4. Heckl D, Kowalczyk MS, Yudovich D, et al. Generation of mouse models of myeloid malignancy with combinatorial genetic lesions using CRISPR-Cas9 genome editing. Nat Biotechnol. 2014; 32(9): 941-946. doi: 10.1038/nbt.2951.
5. Ding DC, Chang YH, Shyu WC, Lin SZ. Human umbilical cord mesenchymal stem cells: a new era for stem cell therapy. Cell Transplant. 2015; 24(3): 339-347. doi: 10.3727/096368915X686841.
6. Pasmant E, Gilbert-Dussardier B, Petit A, et al. SPRED1, a RAS MAPK pathway inhibitor that causes Legius syndrome, is a tumour suppressor downregulated in paediatric acute myeloblastic leukaemia. Oncogene. 2015; 34(5): 631-638. doi: 10.1038/onc.2013.587.
7. Bak RO, Dever DP, Porteus MH. CRISPR/Cas9 genome editing in human hematopoietic stem cells. Nat Protoc. 2018; 13(2): 358-376. doi: 10.1038/nprot.2017.143.
8. Tadokoro Y., Hirao A, et al. Spred1 Safeguards Hematopoietic Homeostasis against Diet-Induced Systemic Stress. Cell Stem Cell. 2018 May 3; 22(5): 713-725.e8. doi: 10.1016/j.stem.2018.04.002. Epub 2018 Apr 26. PMID: 29706577.