An Investigation on Biophysical parameters and Role of Magnesium concentration on spCas9 interaction with Target and Off-Target sequence

Deependra Kumar Ban; Kshama Parate; Deepta Laxmi Bharadwaj; Austin Wong; Lorelai Schoch; Kenneth Visk; Kiana Aran

doi:10.1039/d5sd00227c

January 1, 2025Sensors & DiagnosticsOpen Access

An Investigation on Biophysical parameters and Role of Magnesium concentration on spCas9 interaction with Target and Off-Target sequence

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Key Points

This research aims to understand how magnesium concentration influences the interaction of spCas9 with target and off-target sequences.
Utilized biophysical assays to study spCas9 interactions with DNA sequences
Examined effects of varying magnesium concentrations on specificity
Analyzed binding affinities and recognition patterns of Cas9 to target versus off-target sites
Demonstrated that higher magnesium concentrations enhance spCas9's specificity towards target sequences
Identified altered binding dynamics at single-nucleotide polymorphisms, impacting overall editing efficiency
Showed varying off-target effects correlated with magnesium levels, emphasizing the need for precise control

Implication

This investigation explores magnesium's role in spCas9's target recognition, indicating its significance in genome editing precision.

Abstract

CRISPR-Cas9 enables curative genome editing but requires precise control of target recognition, particularly when single-nucleotide polymorphisms (SNPs) influence specificity. Conventional biochemical and optical assays often rely on endpoint or ensemble-averaged...

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Cite This Study

Ban et al. (2025) studied this question.

synapsesocial.com/papers/69b25aca96eeacc4fcec8e4a https://doi.org/10.1039/d5sd00227c

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An Investigation on Biophysical parameters and Role of Magnesium concentration on spCas9 interaction with Target and Off-Target sequence

Key Points

Implication

Abstract

Discussion

Cite This Study

An Investigation on Biophysical parameters and Role of Magnesium concentration on spCas9 interaction with Target and Off-Target sequence

Key Points

Implication

Abstract

Discussion

Cite This Study

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