The mutagenic chain reaction: A potent drive system for dispersing effector transgenes in insect populations

2016 ◽  
Author(s):  
Ethan Bier
Keyword(s):  
Drive System ◽  
Chain Reaction ◽  
Insect Populations ◽  
Mutagenic Chain Reaction

scholarly journals Modeling the Manipulation of Natural Populations by the Mutagenic Chain Reaction

Genetics ◽  
2015 ◽  
Vol 201 (2) ◽  
pp. 425-431 ◽  
Author(s):  
Robert L. Unckless ◽  
Philipp W. Messer ◽  
Tim Connallon ◽  
Andrew G. Clark
Keyword(s):  
Natural Populations ◽  
Chain Reaction ◽  
Mutagenic Chain Reaction

scholarly journals Evidence for insect transmission of rabbit haemorrhagic disease virus

2002 ◽  
Vol 129 (3) ◽  
pp. 655-663 ◽  
Author(s):  
K. A. McCOLL ◽  
J. C. MERCHANT ◽  
J. HARDY ◽  
B. D. COOKE ◽  
A. ROBINSON ◽  
...  
Keyword(s):  
Laboratory Experiments ◽  
Infectious Virus ◽  
Fold Increase ◽  
Field Trials ◽  
Rabbit Haemorrhagic Disease Virus ◽  
Chain Reaction ◽  
Rabbit Haemorrhagic Disease ◽  
Insect Populations ◽  
Polymerase Chain ◽  
Haemorrhagic Disease

The spread of rabbit haemorrhagic disease (RHD) virus from quarantine on Wardang Island to mainland Australia in 1995 suggested that insects could be potential vectors. Field observations and laboratory experiments were conducted to address aspects of this hypothesis. Firstly, the variation in insect populations on the island during the field trials was examined. There was approximately a 1000-fold increase in the number of bushflies, Musca vetustissima, shortly before the spread of the virus. Secondly, M. vetustissima were tested in the laboratory as potential vectors of RHD virus, and it was demonstrated that disease could be transmitted between rabbits by flies. Finally, 13 of 16 insect samples, collected from Wardang Island and from several sites on the mainland following the spread of virus off the island, were positive for the presence of RHD virus by a specific polymerase chain reaction (PCR). Only one sample contained sufficient infectious virus to kill a susceptible rabbit. These data, combined with previously published information on fly biology, suggested that flies, particularly bushflies, may be involved in the transmission of RHD virus. Other possible routes of spread were not assessed in this study.

scholarly journals A Synthetic Gene Drive System for Local, Reversible Modification and Suppression of Insect Populations

2013 ◽  
Vol 23 (8) ◽  
pp. 671-677 ◽  
Author(s):  
Omar S. Akbari ◽  
Kelly D. Matzen ◽  
John M. Marshall ◽  
Haixia Huang ◽  
Catherine M. Ward ◽  
...  
Keyword(s):  
Drive System ◽  
Synthetic Gene ◽  
Gene Drive ◽  
Insect Populations ◽  
Reversible Modification

scholarly journals Theoretical consequences of the Mutagenic Chain Reaction for manipulating natural populations

2015 ◽  
Author(s):  
Robert Unckless ◽  
Philipp Messer ◽  
Andrew Clark
Keyword(s):  
Critical Point ◽  
High Frequency ◽  
Natural Populations ◽  
Meiotic Drive ◽  
Fitness Cost ◽  
Chain Reaction ◽  
Internal Equilibrium ◽  
Rapid Fixation ◽  
Genetic Technologies ◽  
Mutagenic Chain Reaction

The use of recombinant genetic technologies for population manipulation has mostly remained an abstract idea due to the lack of a suitable means to drive novel gene constructs to high frequency in populations. Recently Gantz and Bier showed that the use of CRISPR/Cas9 technology could provide an artificial drive mechanism, the so-called Mutagenic Chain Reaction (MCR), which could lead to rapid fixation of even a deleterious introduced allele. We establish the equivalence of this system to models of meiotic drive and review the results of simple models showing that, when there is a fitness cost to the MCR allele, an internal equilibrium exists that is usually unstable. Introductions must be at a frequency above this critical point for the successful invasion of the MCR allele. These modeling results have important implications for application of MCR in natural populations.

scholarly journals Autocatalytic–Protection for an Unknown Locus CRISPR–Cas Countermeasure for Undesired Mutagenic Chain Reactions

2020 ◽  
Author(s):  
Ethan Schonfeld ◽  
Elan Schonfeld ◽  
Dan Schonfeld
Keyword(s):  
Genomic Sequence ◽  
Homing Endonuclease ◽  
Gene Drive ◽  
Proof Of Concept ◽  
Chain Reaction ◽  
Gene Encoding ◽  
Chain Reactions ◽  
Homology Directed Repair ◽  
Novel Approach ◽  
Mutagenic Chain Reaction

AbstractThe mutagenic chain reaction (MCR) is a genetic tool to use a CRISPR–Cas construct to introduce a homing endonuclease, allowing gene drive to influence whole populations in a minimal number of generations1,2,3. The question arises: if an active genetic terror event is released into a population, could we prevent the total spread of the undesired allele4? Thus far, MCR protection methods require knowledge of the terror locus5. Here we introduce a novel approach, an autocatalytic-Protection for an Unknown Locus (a-PUL), whose aim is to spread through a population and arrest and decrease an active terror event’s spread without any prior knowledge of the terror-modified locus, thus allowing later natural selection and ERACR drives to restore the normal locus6. a-PUL, using a mutagenic chain reaction, includes (i) a segment encoding a non-Cas9 endonuclease capable of homology-directed repair suggested as Type II endonuclease Cpf1 (Cas12a), (ii) a ubiquitously-expressed gene encoding a gRNA (gRNA1) with a U4AU4 3′-overhang specific to Cpf1 and with crRNA specific to some desired genomic sequence of non-coding DNA, (iii) a ubiquitously-expressed gene encoding two gRNAs (gRNA2/gRNA3) both with tracrRNA specific to Cas9 and crRNA specific to two distinct sites of the Cas9 locus, and (iv) homology arms flanking the Cpf1/gRNA1/gRNA2/gRNA3 cassette that are identical to the region surrounding the target cut directed by gRNA17. We demonstrate the proof-of-concept and efficacy of our protection construct through a Graphical Markov model and computer simulation.

scholarly journals Medusa: A Novel Gene Drive System for Confined Suppression of Insect Populations

PLoS ONE ◽  
2014 ◽  
Vol 9 (7) ◽  
pp. e102694 ◽  
Author(s):  
John M. Marshall ◽  
Bruce A. Hay
Keyword(s):  
Drive System ◽  
Gene Drive ◽  
Novel Gene ◽  
Insect Populations

High performance Nanogold™-in situ hybridzation and its use in the detection of hybridized and PCR-amplified microscopical preparations

1996 ◽  
Vol 54 ◽  
pp. 896-897
Author(s):  
G. W. Hacker ◽  
I. Zehbe ◽  
J. Hainfeld ◽  
A.-H. Graf ◽  
C. Hauser-Kronberger ◽  
...  
Keyword(s):  
Polymerase Chain Reaction ◽  
Messenger Rna ◽  
High Performance ◽  
Detection System ◽  
Direct Methods ◽  
Genetic Alterations ◽  
Chain Reaction ◽  
Protein Encoding ◽  
Polymerase Chain

In situ hybridization (ISH) with biotin-labeled probes is increasingly used in histology, histopathology and molecular biology, to detect genetic nucleic acid sequences of interest, such as viruses, genetic alterations and peptide-/protein-encoding messenger RNA (mRNA). In situ polymerase chain reaction (PCR) (PCR in situ hybridization = PISH) and the new in situ self-sustained sequence replication-based amplification (3SR) method even allow the detection of single copies of DNA or RNA in cytological and histological material. However, there is a number of considerable problems with the in situ PCR methods available today: False positives due to mis-priming of DNA breakdown products contained in several types of cells causing non-specific incorporation of label in direct methods, and re-diffusion artefacts of amplicons into previously negative cells have been observed. To avoid these problems, super-sensitive ISH procedures can be used, and it is well known that the sensitivity and outcome of these methods partially depend on the detection system used.

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