Advancing CRISPR Applications Using Soybean [<i>Glycine max</i> (L.) Merr.] Promoters

Advancing CRISPR Applications Using Soybean [<i>Glycine max</i> (L.) Merr.] Promoters

Bibliographic Details
Main Author: Gunadi, Andika
Language:English
Published: The Ohio State University / OhioLINK 2019
Subjects:
Agriculture
Biology
Botany
Cellular Biology
Molecular Biology
Plant Biology
Plant Sciences
Technology
CRISPR
promoters
particle bombardment
GFP
plant biology
CRISPRi
HITI
genome editing
sgRNA
gene expression
plant biotechnology
Online Access:http://rave.ohiolink.edu/etdc/view?acc_num=osu1566169449003179
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spelling ndltd-OhioLink-oai-etd.ohiolink.edu-osu15661694490031792021-08-03T07:12:25Z Advancing CRISPR Applications Using Soybean [<i>Glycine max</i> (L.) Merr.] Promoters Gunadi, Andika Agriculture Biology Botany Cellular Biology Molecular Biology Plant Biology Plant Sciences Technology CRISPR promoters particle bombardment GFP plant biology CRISPRi HITI genome editing sgRNA gene expression plant biotechnology The advent of clustered regularly interspaced short palindromic repeats (CRISPR) technology has enabled targeted genome modifications in plants through genome editing and gene transcriptional modifications. For high value crops such as soybean [<i>Glycine max</i> (L.) Merr.], CRISPR has the potential to induce targeted genome modifications for crop improvements faster and with higher precision than conventional breeding and transgenic strategies. In its basic form, CRISPR functions through the engineered use of compatible guide RNA and Cas nuclease, which are both simpler and more accessible than presently available alternative targeted genome modification tools such as meganuclease, zinc finger nuclease (ZFN) and transcription activator-like effector nuclease (TALEN). Nevertheless, all of these genome modification tools need to be customized and calibrated for optimal function in plants. A critical customization for CRISPR in soybean is the use of appropriate promoters for introducing the DNA precursors of guide RNA and Cas nuclease. In addition, CRISPR can also be directed to target soybean promoters, leading to precise promoter modifications and modulation of native gene expression. CHAPTER 1 of this dissertation reviews the important functions and applications of promoters in CRISPR-based plant genome modifications. Next, CHAPTER 2 describes the isolation and characterization of 40 unique promoters from soybean, which broaden the toolbox of genetic regulatory components for genome modifications. CHAPTER 3 describes the adaptation of CRISPR interference (CRISPRi) for interrogating <i>cis</i>-elements within a soybean promoter, resulting in pertinent information for future applications of this transcriptional modification strategy. Lastly, CHAPTER 4 describes multiple applications of soybean promoters to both confirm and improve a CRISPR-based targeted DNA integration strategy in plants, termed homology-independent targeted integration (HITI). Customized, particle bombardment-based plant bioassays utilizing the green fluorescence protein marker were developed for testing CRISPRi and HITI efficacies, and have enabled rapid optimizations of both CRISPR approaches. Altogether, the information gleaned from the interconnections of promoters and CRISPR in these studies pave the way towards identifying fundamental mechanisms in plant biology and creative solutions for crop improvements. 2019 English text The Ohio State University / OhioLINK http://rave.ohiolink.edu/etdc/view?acc_num=osu1566169449003179 http://rave.ohiolink.edu/etdc/view?acc_num=osu1566169449003179 unrestricted This thesis or dissertation is protected by copyright: all rights reserved. It may not be copied or redistributed beyond the terms of applicable copyright laws.
collection NDLTD
language English
sources NDLTD
topic Agriculture
Biology
Botany
Cellular Biology
Molecular Biology
Plant Biology
Plant Sciences
Technology
CRISPR
promoters
particle bombardment
GFP
plant biology
CRISPRi
HITI
genome editing
sgRNA
gene expression
plant biotechnology
spellingShingle Agriculture
Biology
Botany
Cellular Biology
Molecular Biology
Plant Biology
Plant Sciences
Technology
CRISPR
promoters
particle bombardment
GFP
plant biology
CRISPRi
HITI
genome editing
sgRNA
gene expression
plant biotechnology
Gunadi, Andika
Advancing CRISPR Applications Using Soybean [<i>Glycine max</i> (L.) Merr.] Promoters
author Gunadi, Andika
author_facet Gunadi, Andika
author_sort Gunadi, Andika
title Advancing CRISPR Applications Using Soybean [<i>Glycine max</i> (L.) Merr.] Promoters
title_short Advancing CRISPR Applications Using Soybean [<i>Glycine max</i> (L.) Merr.] Promoters
title_full Advancing CRISPR Applications Using Soybean [<i>Glycine max</i> (L.) Merr.] Promoters
title_fullStr Advancing CRISPR Applications Using Soybean [<i>Glycine max</i> (L.) Merr.] Promoters
title_full_unstemmed Advancing CRISPR Applications Using Soybean [<i>Glycine max</i> (L.) Merr.] Promoters
title_sort advancing crispr applications using soybean [<i>glycine max</i> (l.) merr.] promoters
publisher The Ohio State University / OhioLINK
publishDate 2019
url http://rave.ohiolink.edu/etdc/view?acc_num=osu1566169449003179
work_keys_str_mv AT gunadiandika advancingcrisprapplicationsusingsoybeaniglycinemaxilmerrpromoters
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