Avoiding Thiol Compound Interference: A Nanoplatform Based on High-Fidelity Au-Se Bonds for Biological Applications

AbstractCRISPR/Cas9 is a programmable genome editing tool that has been widely used for biological applications. While engineered Cas9s have been reported to increase discrimination against off-target cleavage compared with wild type Streptococcus pyogenes (SpCas9) in vivo, the mechanism for enhanced specificity has not been extensively characterized. To understand the basis for improved discrimination against off-target DNA containing important mismatches at the distal end of the guide RNA, we performed kinetic analyses on the high-fidelity (Cas9-HF1) and hyper-accurate (HypaCas9) engineered Cas9 variants. While DNA unwinding is the rate-limiting step for on-target cleavage by SpCas9, we show that chemistry is seriously impaired by more than 100-fold for the high-fidelity variants. The high-fidelity variants improve discrimination by slowing the rate of chemistry without increasing the rate of DNA rewinding—the kinetic partitioning favors release rather than cleavage of a bound off-target substrate because chemistry is slow. Further improvement in discrimination may require engineering increased rates of dissociation of off-target DNA.

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How SIMS microscopy can be used in medicine

Proceedings, annual meeting, Electron Microscopy Society of America ◽

10.1017/s0424820100132649 ◽

1992 ◽

Vol 50 (2) ◽

pp. 1602-1603

Author(s):

Philippe Fragu

Keyword(s):

Mass Spectrometry ◽

Biomedical Applications ◽

Secondary Ion Mass Spectrometry ◽

Chemical Elements ◽

Biological Applications ◽

The Past ◽

Potential Source ◽

Secondary Ion ◽

Chemical Integrity ◽

Scientific Endeavour

The identification, localization and quantification of intracellular chemical elements is an area of scientific endeavour which has not ceased to develop over the past 30 years. Secondary Ion Mass Spectrometry (SIMS) microscopy is widely used for elemental localization problems in geochemistry, metallurgy and electronics. Although the first commercial instruments were available in 1968, biological applications have been gradual as investigators have systematically examined the potential source of artefacts inherent in the method and sought to develop strategies for the analysis of soft biological material with a lateral resolution equivalent to that of the light microscope. In 1992, the prospects offered by this technique are even more encouraging as prototypes of new ion probes appear capable of achieving the ultimate goal, namely the quantitative analysis of micron and submicron regions. The purpose of this review is to underline the requirements for biomedical applications of SIMS microscopy.Sample preparation methodology should preserve both the structural and the chemical integrity of the tissue.

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Using the Theory of Planned Behavior to Predict Faking in Selection Exercises Varying in Fidelity

Journal of Personnel Psychology ◽

10.1027/1866-5888/a000211 ◽

2018 ◽

Vol 17 (3) ◽

pp. 155-160 ◽

Cited By ~ 3

Author(s):

Daniel Dürr ◽

Ute-Christine Klehe

Keyword(s):

Theory Of Planned Behavior ◽

Planned Behavior ◽

Predictive Validity ◽

Group Discussion ◽

Role Play ◽

Selection Procedure ◽

High Fidelity ◽

Selection Research ◽

Play Group

Abstract. Faking has been a concern in selection research for many years. Many studies have examined faking in questionnaires while far less is known about faking in selection exercises with higher fidelity. This study applies the theory of planned behavior (TPB; Ajzen, 1991 ) to low- (interviews) and high-fidelity (role play, group discussion) exercises, testing whether the TPB predicts reported faking behavior. Data from a mock selection procedure suggests that candidates do report to fake in low- and high-fidelity exercises. Additionally, the TPB showed good predictive validity for faking in a low-fidelity exercise, yet not for faking in high-fidelity exercises.

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