Cas9-AAV6-engineered human mesenchymal stromal cells improved cutaneous wound healing in diabetic mice. 2020

Waracharee Srifa, and Nina Kosaric, and Alvaro Amorin, and Othmane Jadi, and Yujin Park, and Sruthi Mantri, and Joab Camarena, and Geoffrey C Gurtner, and Matthew Porteus
Department of Pediatrics, Stanford University School of Medicine, Stanford, CA, 94305, USA.

Human mesenchymal stromal cells (hMSCs) are a promising source for engineered cell-based therapies in which genetic engineering could enhance therapeutic efficacy and install novel cellular functions. Here, we describe an optimized Cas9-AAV6-based genome editing tool platform for site-specific mutagenesis and integration of up to more than 3 kilobases of exogenous DNA in the genome of hMSCs derived from the bone marrow, adipose tissue, and umbilical cord blood without altering their ex vivo characteristics. We generate safe harbor-integrated lines of engineered hMSCs and show that engineered luciferase-expressing hMSCs are transiently active in vivo in wound beds of db/db mice. Moreover, we generate PDGF-BB- and VEGFA-hypersecreting hMSC lines as short-term, local wound healing agents with superior therapeutic efficacy over wildtype hMSCs in the diabetic mouse model without replacing resident cells long-term. This study establishes a precise genetic engineering platform for genetic studies of hMSCs and development of engineered hMSC-based therapies.

UI MeSH Term Description Entries
D007700 Kinetics The rate dynamics in chemical or physical systems.
D002470 Cell Survival The span of viability of a cell characterized by the capacity to perform certain functions such as metabolism, growth, reproduction, some form of responsiveness, and adaptability. Cell Viability,Cell Viabilities,Survival, Cell,Viabilities, Cell,Viability, Cell
D003432 Cross-Linking Reagents Reagents with two reactive groups, usually at opposite ends of the molecule, that are capable of reacting with and thereby forming bridges between side chains of amino acids in proteins; the locations of naturally reactive areas within proteins can thereby be identified; may also be used for other macromolecules, like glycoproteins, nucleic acids, or other. Bifunctional Reagent,Bifunctional Reagents,Cross Linking Reagent,Crosslinking Reagent,Cross Linking Reagents,Crosslinking Reagents,Linking Reagent, Cross,Linking Reagents, Cross,Reagent, Bifunctional,Reagent, Cross Linking,Reagent, Crosslinking,Reagents, Bifunctional,Reagents, Cross Linking,Reagents, Cross-Linking,Reagents, Crosslinking
D003921 Diabetes Mellitus, Experimental Diabetes mellitus induced experimentally by administration of various diabetogenic agents or by PANCREATECTOMY. Alloxan Diabetes,Streptozocin Diabetes,Streptozotocin Diabetes,Experimental Diabetes Mellitus,Diabete, Streptozocin,Diabetes, Alloxan,Diabetes, Streptozocin,Diabetes, Streptozotocin,Streptozocin Diabete
D006801 Humans Members of the species Homo sapiens. Homo sapiens,Man (Taxonomy),Human,Man, Modern,Modern Man
D000072669 Gene Editing Genetic engineering or molecular biology techniques that involve DNA REPAIR mechanisms for incorporating site-specific modifications into a cell's genome. Base Editing,Genome Editing,Editing, Base,Editing, Gene,Editing, Genome
D000076987 CRISPR-Associated Protein 9 An RNA-guided endodeoxyribonuclease that associates with CRISPR SEQUENCES in STREPTOCOCCUS PYOGENES and other bacteria where it participates in an adaptive immune function to cleave foreign DNA complimentary to small GUIDE RNA (sgRNAs). Structurally, Cas9 consists of an ALPHA-HELIX module and a nuclease module connected by a single helix. The nuclease module contains two enzymatic domains: RuvC, which cleaves non-target DNA strand, and an HNH nuclease domain, which cleaves the target strand. Specificity for the DNA target depends on the presence of a protospacer adjacent motif (PAM) sequence, a 2-6 nucleotide DNA sequence immediately following the sequence targeted by Cas9. Cas9 Endonuclease,Cas9 Enzyme,Cas9 Protein,CRISPR Associated Protein 9,Endonuclease, Cas9,Enzyme, Cas9
D000229 Dependovirus A genus of the family PARVOVIRIDAE, subfamily PARVOVIRINAE, which are dependent on a coinfection with helper adenoviruses or herpesviruses for their efficient replication. The type species is Adeno-associated virus 2. Adeno-Associated Viruses,Dependoparvovirus,Adeno-Associated Virus,Virus, Adeno-Associated,Viruses, Adeno-Associated,Adeno Associated Virus,Adeno Associated Viruses,Dependoparvoviruses,Dependoviruses,Virus, Adeno Associated,Viruses, Adeno Associated
D000818 Animals Unicellular or multicellular, heterotrophic organisms, that have sensation and the power of voluntary movement. Under the older five kingdom paradigm, Animalia was one of the kingdoms. Under the modern three domain model, Animalia represents one of the many groups in the domain EUKARYOTA. Animal,Metazoa,Animalia
D012867 Skin The outer covering of the body that protects it from the environment. It is composed of the DERMIS and the EPIDERMIS.

Related Publications

Waracharee Srifa, and Nina Kosaric, and Alvaro Amorin, and Othmane Jadi, and Yujin Park, and Sruthi Mantri, and Joab Camarena, and Geoffrey C Gurtner, and Matthew Porteus
August 2021, Biomaterials,
Waracharee Srifa, and Nina Kosaric, and Alvaro Amorin, and Othmane Jadi, and Yujin Park, and Sruthi Mantri, and Joab Camarena, and Geoffrey C Gurtner, and Matthew Porteus
November 2014, Cytotherapy,
Waracharee Srifa, and Nina Kosaric, and Alvaro Amorin, and Othmane Jadi, and Yujin Park, and Sruthi Mantri, and Joab Camarena, and Geoffrey C Gurtner, and Matthew Porteus
April 2015, Stem cell research & therapy,
Waracharee Srifa, and Nina Kosaric, and Alvaro Amorin, and Othmane Jadi, and Yujin Park, and Sruthi Mantri, and Joab Camarena, and Geoffrey C Gurtner, and Matthew Porteus
May 2023, Diabetes/metabolism research and reviews,
Waracharee Srifa, and Nina Kosaric, and Alvaro Amorin, and Othmane Jadi, and Yujin Park, and Sruthi Mantri, and Joab Camarena, and Geoffrey C Gurtner, and Matthew Porteus
September 2023, Tissue engineering. Part A,
Waracharee Srifa, and Nina Kosaric, and Alvaro Amorin, and Othmane Jadi, and Yujin Park, and Sruthi Mantri, and Joab Camarena, and Geoffrey C Gurtner, and Matthew Porteus
January 2014, Stem cell research & therapy,
Waracharee Srifa, and Nina Kosaric, and Alvaro Amorin, and Othmane Jadi, and Yujin Park, and Sruthi Mantri, and Joab Camarena, and Geoffrey C Gurtner, and Matthew Porteus
June 2017, Journal of dermatological science,
Waracharee Srifa, and Nina Kosaric, and Alvaro Amorin, and Othmane Jadi, and Yujin Park, and Sruthi Mantri, and Joab Camarena, and Geoffrey C Gurtner, and Matthew Porteus
October 2022, International journal of molecular sciences,
Waracharee Srifa, and Nina Kosaric, and Alvaro Amorin, and Othmane Jadi, and Yujin Park, and Sruthi Mantri, and Joab Camarena, and Geoffrey C Gurtner, and Matthew Porteus
January 2020, The Journal of investigative dermatology,
Waracharee Srifa, and Nina Kosaric, and Alvaro Amorin, and Othmane Jadi, and Yujin Park, and Sruthi Mantri, and Joab Camarena, and Geoffrey C Gurtner, and Matthew Porteus
December 2010, Annals of plastic surgery,
Copied contents to your clipboard!