Tuning CRISPR-Cas9 Gene Drives in Saccharomyces cerevisiae

Control of biological populations is an ongoing challenge in many fields, including agriculture, biodiversity, ecological preservation, pest control, and the spread of disease. In some cases, such as insects that harbor human pathogens (e.g., malaria), elimination or reduction of a small number of s...

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Main Authors: Emily Roggenkamp, Rachael M. Giersch, Madison N. Schrock, Emily Turnquist, Megan Halloran, Gregory C. Finnigan
Format: Article
Language:English
Published: Oxford University Press 2018-03-01
Series:G3: Genes, Genomes, Genetics
Subjects:
Online Access:http://g3journal.org/lookup/doi/10.1534/g3.117.300557
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author Emily Roggenkamp
Rachael M. Giersch
Madison N. Schrock
Emily Turnquist
Megan Halloran
Gregory C. Finnigan
author_facet Emily Roggenkamp
Rachael M. Giersch
Madison N. Schrock
Emily Turnquist
Megan Halloran
Gregory C. Finnigan
author_sort Emily Roggenkamp
collection DOAJ
description Control of biological populations is an ongoing challenge in many fields, including agriculture, biodiversity, ecological preservation, pest control, and the spread of disease. In some cases, such as insects that harbor human pathogens (e.g., malaria), elimination or reduction of a small number of species would have a dramatic impact across the globe. Given the recent discovery and development of the CRISPR-Cas9 gene editing technology, a unique arrangement of this system, a nuclease-based “gene drive,” allows for the super-Mendelian spread and forced propagation of a genetic element through a population. Recent studies have demonstrated the ability of a gene drive to rapidly spread within and nearly eliminate insect populations in a laboratory setting. While there are still ongoing technical challenges to design of a more optimal gene drive to be used in wild populations, there are still serious ecological and ethical concerns surrounding the nature of this powerful biological agent. Here, we use budding yeast as a safe and fully contained model system to explore mechanisms that might allow for programmed regulation of gene drive activity. We describe four conserved features of all CRISPR-based drives and demonstrate the ability of each drive component—Cas9 protein level, sgRNA identity, Cas9 nucleocytoplasmic shuttling, and novel Cas9-Cas9 tandem fusions—to modulate drive activity within a population.
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spelling doaj.art-e4f9ead68388442b8f36c6a19c1859932022-12-21T18:23:06ZengOxford University PressG3: Genes, Genomes, Genetics2160-18362018-03-0183999101810.1534/g3.117.30055723Tuning CRISPR-Cas9 Gene Drives in Saccharomyces cerevisiaeEmily RoggenkampRachael M. GierschMadison N. SchrockEmily TurnquistMegan HalloranGregory C. FinniganControl of biological populations is an ongoing challenge in many fields, including agriculture, biodiversity, ecological preservation, pest control, and the spread of disease. In some cases, such as insects that harbor human pathogens (e.g., malaria), elimination or reduction of a small number of species would have a dramatic impact across the globe. Given the recent discovery and development of the CRISPR-Cas9 gene editing technology, a unique arrangement of this system, a nuclease-based “gene drive,” allows for the super-Mendelian spread and forced propagation of a genetic element through a population. Recent studies have demonstrated the ability of a gene drive to rapidly spread within and nearly eliminate insect populations in a laboratory setting. While there are still ongoing technical challenges to design of a more optimal gene drive to be used in wild populations, there are still serious ecological and ethical concerns surrounding the nature of this powerful biological agent. Here, we use budding yeast as a safe and fully contained model system to explore mechanisms that might allow for programmed regulation of gene drive activity. We describe four conserved features of all CRISPR-based drives and demonstrate the ability of each drive component—Cas9 protein level, sgRNA identity, Cas9 nucleocytoplasmic shuttling, and novel Cas9-Cas9 tandem fusions—to modulate drive activity within a population.http://g3journal.org/lookup/doi/10.1534/g3.117.300557CRISPRCas9budding yeastgene drivesgRNAregulating gene drivesbiotechnology
spellingShingle Emily Roggenkamp
Rachael M. Giersch
Madison N. Schrock
Emily Turnquist
Megan Halloran
Gregory C. Finnigan
Tuning CRISPR-Cas9 Gene Drives in Saccharomyces cerevisiae
G3: Genes, Genomes, Genetics
CRISPR
Cas9
budding yeast
gene drive
sgRNA
regulating gene drives
biotechnology
title Tuning CRISPR-Cas9 Gene Drives in Saccharomyces cerevisiae
title_full Tuning CRISPR-Cas9 Gene Drives in Saccharomyces cerevisiae
title_fullStr Tuning CRISPR-Cas9 Gene Drives in Saccharomyces cerevisiae
title_full_unstemmed Tuning CRISPR-Cas9 Gene Drives in Saccharomyces cerevisiae
title_short Tuning CRISPR-Cas9 Gene Drives in Saccharomyces cerevisiae
title_sort tuning crispr cas9 gene drives in saccharomyces cerevisiae
topic CRISPR
Cas9
budding yeast
gene drive
sgRNA
regulating gene drives
biotechnology
url http://g3journal.org/lookup/doi/10.1534/g3.117.300557
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AT madisonnschrock tuningcrisprcas9genedrivesinsaccharomycescerevisiae
AT emilyturnquist tuningcrisprcas9genedrivesinsaccharomycescerevisiae
AT meganhalloran tuningcrisprcas9genedrivesinsaccharomycescerevisiae
AT gregorycfinnigan tuningcrisprcas9genedrivesinsaccharomycescerevisiae