Matching Items (6)
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Description
Extensive efforts have been made to develop efficient and low-cost methods for diagnostics to identify molecular biomarkers that are linked to a wide array of conditions, including cancer. A highly developed method includes utilizing the gene-editing enzyme CRISPR-Cas12a (Cpf1), which demonstrates double-stranded DNase activity with RuvC catalytic domain with high

Extensive efforts have been made to develop efficient and low-cost methods for diagnostics to identify molecular biomarkers that are linked to a wide array of conditions, including cancer. A highly developed method includes utilizing the gene-editing enzyme CRISPR-Cas12a (Cpf1), which demonstrates double-stranded DNase activity with RuvC catalytic domain with high sensitivity and specificity. This DNase activity is RNA-guided and requires a T-rich PAM site on the target sequence for functional cleavage. There have been recent efforts to utilize this DNase activity of Cas12a by combining it with isothermal amplification and analysis by lateral strip tests. This project examined CRISPR-based early detection of microRNA biomarkers. MicroRNA are short RNA molecules that have large roles in post-transcriptional gene regulation. However, due the short length of microRNA and its single-stranded nature, it is challenging to use Cas12a for microRNA detection using existing methods. Thus, this project investigated the potential of two microRNA detection strategies for recognition by CRISPR-Cas12a. These methods were microRNA-splinted ligation with polymerase chain reaction (PCR) and MicroRNA-specific reverse transcriptase PCR (RT-PCR). Gel imaging demonstrated effective amplification of ligated DNA through microRNA-splinted ligation with PCR/RPA. In addition, lateral strips tests showed effective cleavage of the target sequences by Cas12a. However, RT-PCR method demonstrated low amplification by PCR and inefficient poly(A) elongation. This project paves the way for the detection of an extensive range of microRNA biomarkers that are linked to an array of diseases. Future directions include analysis and modifications of RT-PCR method to improve experimental results, extending these detection methods to a larger range of microRNA sequences, and eventually utilizing them for detection in human samples.
ContributorsStaren, Michael Steven (Author) / Green, Alexander (Thesis director) / Stephanopoulos, Nicholas (Committee member) / Diehnelt, Chris (Committee member) / School of Life Sciences (Contributor) / College of Health Solutions (Contributor) / Barrett, The Honors College (Contributor)
Created2019-05
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Description
Advancements in science and technology, particularly in the field of genome editing, hold significant potential to change how future generations will treat disease and may fundamentally change what it means to be human. There are concerns by scientists and non-scientists about how to explore the values and perceptions of the

Advancements in science and technology, particularly in the field of genome editing, hold significant potential to change how future generations will treat disease and may fundamentally change what it means to be human. There are concerns by scientists and non-scientists about how to explore the values and perceptions of the public regarding the implications of new technologies. Use of participatory Technology Assessment (pTA) has arisen as a type of interactive group discussion to disseminate information about technology and collect non-scientists' perceptions of the value, impact or usefulness of a technology and potential ethical issues or consequences to be considered. There is no one size fits all model of pTA; several are discussed in this paper, but there are similarities between them such as the structure of engagement or recruitment criteria. It is important to note a difference in public understanding of science and public engagement with science as it relates to the structure and execution of pTA. This study was undertaken to evaluate pTA as a tool to explore perceptions, values and opinions regarding a case study of CRISPR-Cas9, a tool for genome editing, among ASU Barrett undergraduate students.
ContributorsChapin, Natalie Ann (Author) / Brian, Jennifer (Thesis director) / Bennett, Ira (Committee member) / School of Life Sciences (Contributor) / Department of Psychology (Contributor) / Barrett, The Honors College (Contributor)
Created2016-12
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Description

Multiplex Automated Genome Engineering, or MAGE, is a genome editing technique that enables scientists to quickly edit an organism’s DNA to produce multiple changes across the genome. In 2009, two genetic researchers at the Wyss Institute at Harvard Medical School in Boston, Massachusetts, Harris Wang and George Church, developed the

Multiplex Automated Genome Engineering, or MAGE, is a genome editing technique that enables scientists to quickly edit an organism’s DNA to produce multiple changes across the genome. In 2009, two genetic researchers at the Wyss Institute at Harvard Medical School in Boston, Massachusetts, Harris Wang and George Church, developed the technology during a time when researchers could only edit one site in an organism’s genome at a time. Wang and Church called MAGE a form of accelerated evolution because it creates different cells with many variations of the same original genome over multiple generations. MAGE made genome editing much faster, cheaper, and easier for genetic researchers to create organisms with novel functions that they can use for a variety of purposes, such as making chemicals and medicine, developing biofuels, or further studying and understanding the genes that can cause harmful mutations in humans.

Created2020-12-10
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George McDonald Church studied DNA from living and from extinct species in the US during the twentieth and twenty-first centuries. Church helped to develop and refine techniques with which to describe the complete sequence of all the DNA nucleotides in an organism's genome, techniques such as multiplex sequencing, polony sequencing,

George McDonald Church studied DNA from living and from extinct species in the US during the twentieth and twenty-first centuries. Church helped to develop and refine techniques with which to describe the complete sequence of all the DNA nucleotides in an organism's genome, techniques such as multiplex sequencing, polony sequencing, and nanopore sequencing. Church also contributed to the Human Genome Project, and in 2005 he helped start a company, the Personal Genome Project. Church proposed to use DNA from extinct species to clone and breed new organisms from those species.

Created2015-08-12
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Description

In 2018, He Jiankui uploaded a series of videos to a YouTube channel titled “The He Lab” that detailed one of the first instances of a successful human birth after genome editing had been performed on an embryo using CRISPR-cas9. CRISPR-cas9 is a genome editing tool derived from bacteria that

In 2018, He Jiankui uploaded a series of videos to a YouTube channel titled “The He Lab” that detailed one of the first instances of a successful human birth after genome editing had been performed on an embryo using CRISPR-cas9. CRISPR-cas9 is a genome editing tool derived from bacteria that can be used to cut out and replace specific sequences of DNA. He genetically modified embryos at his lab in Shenzhen, China, to make them immune to contracting HIV through indirect perinatal transmission from their father, who was infected with the virus. HIV is a virus that attacks the immune cells of its host and weakens their ability to fight off diseases. At the time of He’s experiment, various treatments already existed at that could prevent the fetuses from contracting HIV without the need for gene surgery. Nonetheless, He’s experiment led to one of the first successful births of fetuses resulting from genetically modified embryos. He kept his experiment secret until he uploaded the videos announcing the birth of the fetuses, born as two twin girls. The experiment discussed in the videos was successful, but many scientists criticized the experiment due to ethical concerns with the way He conducted it.

Created2021-07-31