scholarly journals Selection-Free Non-Viral Method Revealed Highly Efficient Crispr-Cas9 Genome Editing of Human Pluripotent Stem Cells Guided by Cellular Autophagy

2021 ◽  
Author(s):  
Michelle Surma ◽  
Kavitha Anbarasu ◽  
Arupratan Das
Keyword(s):  
Stem Cells ◽  
Genome Editing ◽  
Pluripotent Stem Cells ◽  
Human Pluripotent Stem Cells ◽  
Highly Efficient ◽  
Cellular Autophagy

scholarly journals Highly Efficient CRISPR-Cas9-Mediated Genome Editing in Human Pluripotent Stem Cells

2018 ◽  
Vol 48 (1) ◽  
pp. e64 ◽  
Author(s):  
Jean Ann Maguire ◽  
Fabian L. Cardenas-Diaz ◽  
Paul Gadue ◽  
Deborah L. French
Keyword(s):  
Stem Cells ◽  
Genome Editing ◽  
Pluripotent Stem Cells ◽  
Human Pluripotent Stem Cells ◽  
Highly Efficient

scholarly journals Highly Efficient and Marker-free Genome Editing of Human Pluripotent Stem Cells by CRISPR-Cas9 RNP and AAV6 Donor-Mediated Homologous Recombination

2019 ◽  
Vol 24 (5) ◽  
pp. 821-828.e5 ◽  
Author(s):  
Renata M. Martin ◽  
Kazuya Ikeda ◽  
M. Kyle Cromer ◽  
Nobuko Uchida ◽  
Toshinobu Nishimura ◽  
...  
Keyword(s):  
Stem Cells ◽  
Homologous Recombination ◽  
Genome Editing ◽  
Pluripotent Stem Cells ◽  
Human Pluripotent Stem Cells ◽  
Highly Efficient ◽  
Marker Free

scholarly journals Highly efficient genome editing via CRISPR–Cas9 in human pluripotent stem cells is achieved by transient BCL-XL overexpression

2018 ◽  
Vol 46 (19) ◽  
pp. 10195-10215 ◽  
Author(s):  
Xiao-Lan Li ◽  
Guo-Hua Li ◽  
Juan Fu ◽  
Ya-Wen Fu ◽  
Lu Zhang ◽  
...  
Keyword(s):  
Stem Cells ◽  
Genome Editing ◽  
Pluripotent Stem Cells ◽  
Human Pluripotent Stem Cells ◽  
Highly Efficient

scholarly journals Selection-free non-viral method revealed highly efficient CRISPR-Cas9 genome editing of human pluripotent stem cells guided by cellular autophagy

2021 ◽  
Author(s):  
Michelle Surma ◽  
Kavitha Anbarasu ◽  
Arupratan Das
Keyword(s):  
Stem Cells ◽  
Genome Editing ◽  
Pluripotent Stem Cells ◽  
Disease Modeling ◽  
Human Pluripotent Stem Cells ◽  
Cell Replacement Therapy ◽  
Gene Expressions ◽  
Guide Rna ◽  
Transfection Method ◽  
Cellular Autophagy

CRISPR-Cas9 mediated genome editing of human pluripotent stem cells (hPSCs) provides strong avenues for human disease modeling, drug discovery and cell replacement therapy. Genome editing of hPSCs is an extremely inefficient process and requires complex gene delivery and selection methods to improve edit efficiency which are not ideal for clinical applications. Here, we have shown a selection free simple lipofectamine based transfection method where a single plasmid encoding guide RNA (gRNA) and Cas9 selectively transfected hPSCs at the colony edges. Upon dissection and sequencing, the edge cells showed more than 30% edit frequency compared to the reported 3% rate under no selections. Increased cellular health of the edge cells as revealed by reduced autophagy gene-expressions is critical for such transfection pattern. Edge specific transfection was inhibited by blocking lysosomal activity which is essential for autophagy. Hence, our method provides robust scarless genome-editing of hPSCs which is ideal for translational research.

scholarly journals An iCRISPR Platform for Rapid, Multiplexable, and Inducible Genome Editing in Human Pluripotent Stem Cells

2014 ◽  
Vol 15 (2) ◽  
pp. 215-226 ◽  
Author(s):  
Federico González ◽  
Zengrong Zhu ◽  
Zhong-Dong Shi ◽  
Katherine Lelli ◽  
Nipun Verma ◽  
...  
Keyword(s):  
Stem Cells ◽  
Genome Editing ◽  
Pluripotent Stem Cells ◽  
Human Pluripotent Stem Cells

scholarly journals Genome Editing Human Pluripotent Stem Cells to Model β-Cell Disease and Unmask Novel Genetic Modifiers

2021 ◽  
Vol 12 ◽  
Author(s):  
Matthew N. George ◽  
Karla F. Leavens ◽  
Paul Gadue
Keyword(s):  
Stem Cells ◽  
Genome Editing ◽  
Pluripotent Stem Cells ◽  
Genetic Disease ◽  
Single Gene ◽  
Monogenic Diabetes ◽  
Human Pluripotent Stem Cells ◽  
Genetic Modifiers ◽  
Β Cell ◽  
Protein Coding

A mechanistic understanding of the genetic basis of complex diseases such as diabetes mellitus remain elusive due in large part to the activity of genetic disease modifiers that impact the penetrance and/or presentation of disease phenotypes. In the face of such complexity, rare forms of diabetes that result from single-gene mutations (monogenic diabetes) can be used to model the contribution of individual genetic factors to pancreatic β-cell dysfunction and the breakdown of glucose homeostasis. Here we review the contribution of protein coding and non-protein coding genetic disease modifiers to the pathogenesis of diabetes subtypes, as well as how recent technological advances in the generation, differentiation, and genome editing of human pluripotent stem cells (hPSC) enable the development of cell-based disease models. Finally, we describe a disease modifier discovery platform that utilizes these technologies to identify novel genetic modifiers using induced pluripotent stem cells (iPSC) derived from patients with monogenic diabetes caused by heterozygous mutations.

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