scholarly journals Application of CRISPR/Cas9 to human-induced pluripotent stem cells: from gene editing to drug discovery

2020 ◽  
Vol 14 (1) ◽  
Author(s):  
Claudia De Masi ◽  
Paola Spitalieri ◽  
Michela Murdocca ◽  
Giuseppe Novelli ◽  
Federica Sangiuolo
2016 ◽  
Vol 17 (2) ◽  
pp. 256 ◽  
Author(s):  
Mohammed Kawser Hossain ◽  
Ahmed Abdal Dayem ◽  
Jihae Han ◽  
Subbroto Kumar Saha ◽  
Gwang-Mo Yang ◽  
...  

2020 ◽  
Author(s):  
Engi Ahmed ◽  
Mathieu Fieldes ◽  
Chloé Bourguignon ◽  
Joffrey Mianné ◽  
Aurélie Petit ◽  
...  

AbstractRationaleHighly reproducible in vitro generation of human bronchial epithelium from pluripotent stem cells is an unmet key goal for drug screening to treat lung diseases. The possibility of using induced pluripotent stem cells (hiPSC) to model normal and diseased tissue in vitro from a simple blood sample will reshape drug discovery for chronic lung, monogenic and infectious diseases.MethodsWe devised a simple and reliable method that drives a blood sample reprogrammed into hiPSC subsequently differentiated within 45 days into air-liquid interface bronchial epithelium (iALI), through key developmental stages, definitive-endoderm (DE) and Ventralized-Anterior-Foregut-Endoderm (vAFE) cells.ResultsReprogramming blood cells from one healthy and 3 COPD patients, and from skin-derived fibroblasts obtained in one PCD patient, succeeded in 100% of samples using Sendai viruses. Mean cell purity at DE and vAFE stages was greater than 80%, assessed by expression of CXCR4 and NKX2.1, avoiding the need of cell sorting. When transferred to ALI conditions, vAFE cells reliably differentiated within 4 weeks into bronchial epithelium with large zones covered by beating ciliated, basal, goblets, club cells and neuroendocrine cells as found in vivo. Benchmarking all culture conditions including hiPSCs adaptation to single-cell passaging, cell density and differentiation induction timing allowed for consistently producing iALI bronchial epithelium from the five hiPSC lines.ConclusionsReliable reprogramming and differentiation of blood-derived hiPSCs into mature and functional iALI bronchial epithelium is ready for wider use and this will allow better understanding lung disease pathogenesis and accelerating the development of novel gene therapies and drug discovery.


2012 ◽  
Vol 120 (1) ◽  
pp. 103-111 ◽  
Author(s):  
Lezanne Ooi ◽  
Kuldip Sidhu ◽  
Anne Poljak ◽  
Greg Sutherland ◽  
Michael D. O’Connor ◽  
...  

Methods ◽  
2017 ◽  
Vol 121-122 ◽  
pp. 29-44 ◽  
Author(s):  
Saniye Yumlu ◽  
Jürgen Stumm ◽  
Sanum Bashir ◽  
Anne-Kathrin Dreyer ◽  
Pawel Lisowski ◽  
...  

Cells ◽  
2021 ◽  
Vol 10 (10) ◽  
pp. 2698
Author(s):  
Ishnoor Sidhu ◽  
Sonali P. Barwe ◽  
Raju K. Pillai ◽  
Anilkumar Gopalakrishnapillai

In vitro modeling of hematological malignancies not only provides insights into the influence of genetic aberrations on cellular and molecular mechanisms involved in disease progression but also aids development and evaluation of therapeutic agents. Owing to their self-renewal and differentiation capacity, induced pluripotent stem cells (iPSCs) have emerged as a potential source of short in supply disease-specific human cells of the hematopoietic lineage. Patient-derived iPSCs can recapitulate the disease severity and spectrum of prognosis dictated by the genetic variation among patients and can be used for drug screening and studying clonal evolution. However, this approach lacks the ability to model the early phases of the disease leading to cancer. The advent of genetic editing technology has promoted the generation of precise isogenic iPSC disease models to address questions regarding the underlying genetic mechanism of disease initiation and progression. In this review, we discuss the use of iPSC disease modeling in hematological diseases, where there is lack of patient sample availability and/or difficulty of engraftment to generate animal models. Furthermore, we describe the power of combining iPSC and precise gene editing to elucidate the underlying mechanism of initiation and progression of various hematological malignancies. Finally, we discuss the power of iPSC disease modeling in developing and testing novel therapies in a high throughput setting.


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