Chain Reaction Systems Based on Loop Dissociation of DNA

2006 ◽  
pp. 347-358 ◽  
Author(s):  
Keiichiro Takahashi ◽  
Satsuki Yaegashi ◽  
Atsushi Kameda ◽  
Masami Hagiya
Keyword(s):  
Chain Reaction ◽  
Reaction Systems

scholarly journals CRISPR/Cas9-mediated base-editing enables a chain reaction through sequential repair of sgRNA scaffold mutations

2021 ◽  
Vol 11 (1) ◽  
Author(s):  
Tsuyoshi Fukushima ◽  
Yosuke Tanaka ◽  
Keito Adachi ◽  
Nanami Masuyama ◽  
Akiho Tsuchiya ◽  
...  
Keyword(s):  
Mammalian Cells ◽  
Regulatory Networks ◽  
Chain Reaction ◽  
Reaction Systems ◽  
Chain Reactions ◽  
Base Editing ◽  
A Chain ◽  
Loxp Sequence

AbstractCell behavior is controlled by complex gene regulatory networks. Although studies have uncovered diverse roles of individual genes, it has been challenging to record or control sequential genetic events in living cells. In this study, we designed two cellular chain reaction systems that enable sequential sgRNA activation in mammalian cells using a nickase Cas9 tethering of a cytosine nucleotide deaminase (nCas9-CDA). In these systems, thymidine (T)-to-cytosine (C) substitutions in the scaffold region of the sgRNA or the TATA box-containing loxP sequence (TATAloxP) are corrected by the nCas9-CDA, leading to activation of the next sgRNA. These reactions can occur multiple times, resulting in cellular chain reactions. As a proof of concept, we established a chain reaction by repairing sgRNA scaffold mutations in 293 T cells. Importantly, the results obtained in yeast or in vitro did not match those obtained in mammalian cells, suggesting that in vivo chain reactions need to be optimized in appropriate cellular contexts. Our system may lay the foundation for building cellular chain reaction systems that have a broad utility in the future biomedical research.

Use of Single- and Multi-Locus and Polymerase Chain Reaction Systems for Zygosity Determination - Clinical Application in Twins with Clefts of the Lip and Palate

1995 ◽  
Vol 44 (1) ◽  
pp. 25-30 ◽  
Author(s):  
H. Eufinger ◽  
S.P. Rand ◽  
U. Schütte
Keyword(s):  
Polymerase Chain Reaction ◽  
Clinical Application ◽  
Dna Analysis ◽  
Chain Reaction ◽  
Probability Of Error ◽  
Oral Clefts ◽  
Reaction Systems ◽  
Dna Sampling ◽  
Modern Methods ◽  
Polymerase Chain

AbstractPrecision of zygosity determination in twins can be improved by the use of modern methods of DNA analysis. The clinical application of 4 single- (SLS) and 2 multi-locus (MLS), and 6 PCR (polymerase chain reaction) systems for zygosity determination in 12 twin pairs with oral clefts was compared with regard to the quality and quantity of sample material required and the probability of error in monozygosity determination. PCR systems proved to be superior to SLS or MLS, as DNA sampling is much more convenient, while its level of accuracy still fulfils clinical requirements. For this reason, PCR systems should be considered a basic method in modern clinical twin research.

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