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http://dx.doi.org/10.1186/s12866-021-02100-w | DOI Listing |
Gaucher disease type 1 is a lysosomal storage disorder caused by mutations that reduce glucocerebrosidase activity, leading to glycolipid buildup, particularly in macrophages. To develop a curative approach, we established a high-efficiency genome editing platform for human and murine hematopoietic stem-progenitor cells using CRISPR/Cas9, recombinant adeno-associated virus serotype 6. To enhance homology-directed DNA repair while minimizing genotoxicity, we incorporated a new 53BP1 inhibitor, a ubiquitin variant that promotes DNA end resection and significantly increases editing efficiency.
View Article and Find Full Text PDFBio Protoc
August 2025
Institute of Health Sciences and Technology (IHST), Institutes of Physical Sciences and Information Technology, Anhui University, Hefei, China.
The Sox (SRY-related HMG-box) protein family plays a crucial role in cellular differentiation, development, and gene regulation, with the HMG (high-mobility group) domain responsible for DNA binding and transcriptional regulation. Proteins in the SOX gene family contain an HMG domain that shares 50% homology with the HMG domain of the sex-determining factor SRY gene. The SOX gene family comprises 30 proteins, which are classified into 10 groups (A-H).
View Article and Find Full Text PDFMol Ther Methods Clin Dev
September 2025
Division of Translational Science and Therapeutics, Fred Hutchinson Cancer Center, Seattle, WA 98109, USA.
Autologous transplantation of gene-modified/corrected hematopoietic stem cells (HSCs) offers a definitive therapeutic approach to restore hematopoiesis in Fanconi anemia (FA) patients. However, this approach not only requires treatment of patient HSCs in specialized facilities but also inevitably results in a loss of fragile and limited patient HSCs. correction of HSCs directly in the patient can overcome these limitations.
View Article and Find Full Text PDFNucleic Acids Res
August 2025
Department of Biomedicine, Aarhus University, Aarhus C 8000, Denmark.
Repair of double-strand DNA breaks generated by site-directed endonucleases, like Cas9, is the hallmark of gene editing based on homology-directed repair (HDR). HDR uses an exogenous DNA template to restore the cleaved DNA sequence and can facilitate specific gene corrections as well as insertion of genes or partial complementary DNA (cDNA) sequences. For CRISPR/Cas-directed gene editing, co-administration of the Cas9/single guide RNA (sgRNA) ribonucleoprotein (RNP) complex and a DNA template typically involves two different delivery strategies or different types of vehicles.
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