Off Target Effects
Off Target Effects - Related Videos
11:37
Using Sniper-Cas9 to Minimize Off-target Effects of CRISPR-Cas9 Without the Loss of On-target Activity Via Directed Evolution
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Cited by 21 •
2019
Here, we present a protocol to optimize CRISPR-Cas9 to achieve a higher specificity without the loss of on-target activity. We use a directed evolution approach called Sniper-screen to find a mutant Cas9 with the desired characteristics. Sniper-Cas9 is compatible with truncated single-guide RNAs and delivery in a ribonucleoprotein format, well-known strategies for achieving higher specificities.
09:04
Comprehensive Evaluation of the Effectiveness and Safety of Placenta-Targeted Drug Delivery Using Three Complementary Methods
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Cited by 10 •
2018
We describe a system that utilizes three methods to evaluate the safety and effectiveness of placenta-targeted drug delivery: in vivo imaging to monitor nanoparticle accumulation, high-frequency ultrasound to monitor placental and fetal development, and HPLC to quantify drug delivery to tissue.
08:56
Recombineering and Gene Targeting
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2023
One of the most widely used tools in modern biology is molecular cloning with restriction enzymes, which create compatible ends between DNA fragments that allow them to be joined together. However, this technique has certain restrictions that limit its applicability for large or complex DNA construct generation. A newer technique that addresses some of these shortcomings is recombineering, which modifies DNA using homologous recombination (HR), the exchange between different DNA molecules based...
02:19
Buffer Effectiveness
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2020
Buffer solutions do not have an unlimited capacity to keep the pH relatively constant . Instead, the ability of a buffer solution to resist changes in pH relies on the presence of appreciable amounts of its conjugate weak acid-base pair. When enough strong acid or base is added to substantially lower the concentration of either member of the buffer pair, the buffering action within the solution is compromised. The buffer capacity is the amount of acid or base that can be added to a given volume...
06:03
Operational and Intervention Effects of Targeted Tuina in Lumbar Intervertebral Disc Degeneration Model Rabbits
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Cited by 1 •
2023
The finger tactile measurement system quantifies tactile pressure on the finger using a highly sensitive electrosurgical pressure sensor, with software displaying and accurately recording the pressure data and video in real time. Two analysis modules unify the data processing for thumb tracking and conditioning during Tuina.
FAQs
Several molecular routes can produce Off Target Effects, and each points to a different strategy for improving specificity. Nonspecific binding may cause an intervention to interact with unrelated molecules or pathways, while structural similarity can make unintended targets susceptible. In gene-targeting systems, partial sequence complementarity can redirect activity. Excessive exposure can further increase unintended interactions.
Excessive exposure increases the opportunity for an intervention to act beyond its intended target, so observed biological changes may not reflect the intended mechanism alone. This matters when interpreting experiments or assessing safety because unintended activity can alter cells or pathways and may contribute to toxicity. Evaluating exposure alongside target specificity helps distinguish desired effects from broader biological disruption.
Gene-targeting systems can produce unintended activity through partial sequence complementarity, allowing the intervention to affect sequences beyond the intended one. Other drugs or experimental manipulations may instead act through nonspecific binding, structural similarity between targets, or excessive exposure. Recognizing the mechanism helps researchers select appropriate tests and refine the intervention according to the source of unintended activity.
Computational predictions provide an initial way to identify possible unintended interactions before or alongside experimental testing. They can highlight candidates suggested by structural similarity or partial sequence complementarity, helping researchers focus biochemical, cell-based, or organismal studies. This approach supports more efficient evaluation of target specificity, although the source identifies it as one component of a broader detection strategy.
Biochemical assays can examine whether an intervention acts on molecules beyond its intended target, whereas cell-based tests show whether those interactions produce changes in living cells or pathways. Using both levels connects molecular activity with biological consequences. Their results help researchers interpret experimental findings, evaluate specificity, identify potential toxicity, and guide improvements to the intervention.
Organismal studies extend evaluation beyond isolated molecules or cells to determine whether unintended activity appears in a more complete biological setting. They are relevant when researchers need evidence for safety, toxicity, or the reliability of a therapy or experimental tool. Combined with computational, biochemical, and cell-based approaches, these studies support a broader assessment of biological consequences.