New application of the CRISPR‐Cas9 system for site‐specific exogenous gene doping analysis

2020 ◽  
Author(s):  
Joon‐Yeop Yi ◽  
Minyoung Kim ◽  
Hophil Min ◽  
Byung‐Gee Kim ◽  
Junghyun Son ◽  
...  
Keyword(s):  
Doping Analysis ◽  
Site Specific ◽  
Gene Doping ◽  
Exogenous Gene

Detection of exogenous gene doping of IGF-I by a real-time quantitative PCR assay

2017 ◽  
Vol 64 (4) ◽  
pp. 549-554 ◽  
Author(s):  
Jin-Ju Zhang ◽  
Jing-Feng Xu ◽  
Yong-Wei Shen ◽  
Shi-Jiao Ma ◽  
Ting-Ting Zhang ◽  
...  
Keyword(s):  
Real Time ◽  
Quantitative Pcr ◽  
Pcr Assay ◽  
Gene Doping ◽  
Real Time Quantitative Pcr ◽  
Igf I ◽  
Exogenous Gene ◽  
Quantitative Pcr Assay

Development of a dCas9-Based High-Throughput Gene Doping Analysis (HiGDA) for Human Erythropoietin

2021 ◽  
Author(s):  
Joon-Yeop Yi ◽  
Minyoung Kim ◽  
Byung Gee Kim ◽  
Junghyun Son ◽  
Changmin Sung
Keyword(s):  
High Throughput ◽  
Doping Analysis ◽  
Gene Doping ◽  
Human Erythropoietin

Transgene traceability in transgenic mice: a bioanalytical approach for potential gene-doping analysis

Bioanalysis ◽  
2011 ◽  
Vol 3 (22) ◽  
pp. 2523-2531 ◽  
Author(s):  
Patrizia Bogani ◽  
Maria Michela Spiriti ◽  
Stefano Lazzarano ◽  
Annarosa Arcangeli ◽  
Marcello Buiatti ◽  
...  
Keyword(s):  
Transgenic Mice ◽  
Doping Analysis ◽  
Gene Doping

Large-cluster and combined fluorescent and gold immunoprobes

1996 ◽  
Vol 54 ◽  
pp. 892-893
Author(s):  
Richard D. Powell ◽  
James F. Hainfeld ◽  
Carol M. R. Halsey ◽  
David L. Spector ◽  
Shelley Kaurin ◽  
...  
Keyword(s):  
Metal Cluster ◽  
Sulfonyl Chloride ◽  
Gel Filtration ◽  
Cross Linking ◽  
Site Specific ◽  
Fab Fragments ◽  
Cluster Label ◽  
Covalently Linked ◽  
Texas Red ◽  
Fold Excess

Two new types of covalently linked, site-specific immunoprobes have been prepared using metal cluster labels, and used to stain components of cells. Combined fluorescein and 1.4 nm “Nanogold” labels were prepared by using the fluorescein-conjugated tris (aryl) phosphine ligand and the amino-substituted ligand in the synthesis of the Nanogold cluster. This cluster label was activated by reaction with a 60-fold excess of (sulfo-Succinimidyl-4-N-maleiniido-cyclohexane-l-carboxylate (sulfo-SMCC) at pH 7.5, separated from excess cross-linking reagent by gel filtration, and mixed in ten-fold excess with Goat Fab’ fragments against mouse IgG (obtained by reduction of F(ab’)2 fragments with 50 mM mercaptoethylamine hydrochloride). Labeled Fab’ fragments were isolated by gel filtration HPLC (Superose-12, Pharmacia). A combined Nanogold and Texas Red label was also prepared, using a Nanogold cluster derivatized with both and its protected analog: the cluster was reacted with an eight-fold excess of Texas Red sulfonyl chloride at pH 9.0, separated from excess Texas Red by gel filtration, then deprotected with HC1 in methanol to yield the amino-substituted label.

The challenge of detecting modifications on proteins

2020 ◽  
Vol 64 (1) ◽  
pp. 135-153 ◽  
Author(s):  
Lauren Elizabeth Smith ◽  
Adelina Rogowska-Wrzesinska
Keyword(s):  
Mass Spectrometry ◽  
Protein Function ◽  
Chemical Properties ◽  
Lysine Acetylation ◽  
Mass Determination ◽  
Post Translational Modifications ◽  
Site Specific ◽  
The Common

Abstract Post-translational modifications (PTMs) are integral to the regulation of protein function, characterising their role in this process is vital to understanding how cells work in both healthy and diseased states. Mass spectrometry (MS) facilitates the mass determination and sequencing of peptides, and thereby also the detection of site-specific PTMs. However, numerous challenges in this field continue to persist. The diverse chemical properties, low abundance, labile nature and instability of many PTMs, in combination with the more practical issues of compatibility with MS and bioinformatics challenges, contribute to the arduous nature of their analysis. In this review, we present an overview of the established MS-based approaches for analysing PTMs and the common complications associated with their investigation, including examples of specific challenges focusing on phosphorylation, lysine acetylation and redox modifications.

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