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Full-Text Articles in Engineering
Advances In Homology Directed Genetic Engineering Of Human Pluripotent And Adult Stem Cells, Kalpith Ramamoorthi, Donald Curtis, Prashanth Asuri
Advances In Homology Directed Genetic Engineering Of Human Pluripotent And Adult Stem Cells, Kalpith Ramamoorthi, Donald Curtis, Prashanth Asuri
Bioengineering
The ability to introduce precise genomic modifications in human cells has profound implications for both basic and applied research in stem cells, ranging from identification of genes regulating stem cell self-renewal and multilineage differentiation to therapeutic gene correction and creation of in vitro models of human diseases. However, the overall efficiency of this process is challenged by several factors including inefficient gene delivery into stem cells and low rates of homology directed site-specific targeting. Recent studies report the development of novel techniques to improve gene targeting efficiencies in human stem cells; these methods include molecular engineering of viral vectors o …
Soft Microenvironments Promote The Early Neurogenic Differentiation But Not Self-Renewal Of Human Pluripotent Stem Cells, Albert Keung, Prashanth Asuri, Sanjay Kumar, David Schaffer
Soft Microenvironments Promote The Early Neurogenic Differentiation But Not Self-Renewal Of Human Pluripotent Stem Cells, Albert Keung, Prashanth Asuri, Sanjay Kumar, David Schaffer
Bioengineering
Human pluripotent stem cells (hPSCs) are of great interest in biology and medicine due to their ability to self-renew and differentiate into any adult or fetal cell type. Important efforts have identified biochemical factors, signaling pathways, and transcriptional networks that regulate hPSC biology. However, recent work investigating the effect of biophysical cues on mammalian cells and adult stem cells suggests that the mechanical properties of the microenvironment, such as stiffness, may also regulate hPSC behavior. While several studies have explored this mechanoregulation in mouse embryonic stem cells (mESCs), it has been challenging to extrapolate these findings and thereby explore their …
Directed Evolution Of Adeno-Associated Virus For Enhanced Gene Delivery And Gene Targeting In Human Pluripotent Stem Cells, Prashanth Asuri, Melissa Bartel, Tandis Vazin, Jae-Hyung Jang, Tiffany Wong, David Schaffer
Directed Evolution Of Adeno-Associated Virus For Enhanced Gene Delivery And Gene Targeting In Human Pluripotent Stem Cells, Prashanth Asuri, Melissa Bartel, Tandis Vazin, Jae-Hyung Jang, Tiffany Wong, David Schaffer
Bioengineering
Efficient approaches for the precise genetic engineering of human pluripotent stem cells (hPSCs) can enhance both basic and applied stem cell research. Adenoassociated virus (AAV) vectors are of particular interest for their capacity to mediate efficient gene delivery to and gene targeting in various cells. However, natural AAV serotypes offer only modest transduction of human embryonic and induced pluripotent stem cells (hESCs and hiPSCs), which limits their utility for efficiently manipulating the hPSC genome. Directed evolution is a powerful means to generate viral vectors with novel capabilities, and we have applied this approach to create a novel AAV variant with …
An Evolved Adeno-Associated Viral Variant Enhances Gene Delivery And Gene Targeting In Neural Stem Cells, Jae-Hyung Jang, James Koerber, Jung-Suk Kim, Prashanth Asuri, Tandis Vazin, Melissa Bartel, Albert Keung, Inchan Kwan, David Schaffer
An Evolved Adeno-Associated Viral Variant Enhances Gene Delivery And Gene Targeting In Neural Stem Cells, Jae-Hyung Jang, James Koerber, Jung-Suk Kim, Prashanth Asuri, Tandis Vazin, Melissa Bartel, Albert Keung, Inchan Kwan, David Schaffer
Bioengineering
Gene delivery to, and gene targeting in, stem cells would be a highly enabling technology for basic science and biomedical application. Adeno-associated viral (AAV) vectors have demonstrated the capacity for efficient delivery to numerous cells, but their application to stem cells has been limited by low transduction efficiency. Due to their considerable advantages, however, engineering AAV delivery systems to enhance gene delivery to stem cells may have an impact in stem cell biology and therapy. Therefore, using several diverse AAV capsid libraries—including randomly mutagenized, DNA shuffled, and random peptide insertion variants—we applied directed evolution to create a “designer” AAV vector …