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Related Concept Videos

Genomics02:02

Genomics

40.7K
Genomics is the science of genomes: it is the study of all the genetic material of an organism. In humans, the genome consists of information carried in 23 pairs of chromosomes in the nucleus, as well as mitochondrial DNA. In genomics, both coding and non-coding DNA is sequenced and analyzed. Genomics allows a better understanding of all living things, their evolution, and their diversity. It has a myriad of uses: for example, to build phylogenetic trees, to improve productivity and...
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What is Genetic Engineering?00:49

What is Genetic Engineering?

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Overview
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Spin–Spin Coupling: Two-Bond Coupling (Geminal Coupling)01:20

Spin–Spin Coupling: Two-Bond Coupling (Geminal Coupling)

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Two NMR-active nuclei bonded to a central atom can be involved in geminal or two-bond coupling. Geminal coupling is commonly seen between diastereotopic protons in chiral molecules and unsymmetrical alkenes, among others.
The central atom need not be NMR-active because its electrons are affected by the electron polarization of the spin-active atoms. However, spin information is transmitted less effectively than in one-bond coupling, and 2J values are usually weaker than 1J values. The energy of...
1.7K
Spin–Spin Coupling: Three-Bond Coupling (Vicinal Coupling)01:22

Spin–Spin Coupling: Three-Bond Coupling (Vicinal Coupling)

1.5K
Vicinal or three-bond coupling is commonly observed between protons attached to adjacent carbons. Here, nuclear spin information is primarily transferred via electron spin interactions between adjacent C‑H bond orbitals. This generally favors the antiparallel arrangement of spins, so 3J values are usually positive.
The extent of coupling depends on the C‑C bond length, the two H‑C‑C angles, any electron-withdrawing substituents, and the dihedral angle between the involved orbitals. The...
1.5K
Genomic Imprinting and Inheritance02:30

Genomic Imprinting and Inheritance

37.2K
Diploid organisms inherit genetic material through chromosomes from both parents. Copies of the same gene are known as alleles. In most cases, both alleles are simultaneously expressed and allow various cellular processes to function optimally. If one of the alleles is missing or mutated, the expression of the other allele can compensate; however, this is not true for all genes.
The expression of some genes depends on which parent passed the gene to the offspring, through a phenomenon known as...
37.2K
Data Reporting and Recording01:24

Data Reporting and Recording

5.5K
Reporting and recording are crucial in data documentation. The timely, thorough, and accurate documentation of facts is essential when recording patient data. Failure to record findings during an assessment or interpretation of a problem will result in loss of information and make the patient document unreliable. The reader is left with general impressions if the information is not specific. A recording is documenting data of the individual's health information in a traceable, secure, and...
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Related Experiment Video

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Genome Editing with CompoZr Custom Zinc Finger Nucleases ZFNs
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Genome Editing with CompoZr Custom Zinc Finger Nucleases ZFNs

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An Improved Genome Engineering Method Using Surrogate Reporter-Coupled Suicidal ZFNs.

Jiani Xing1, Cunfang Zhang1, Kun Xu1

  • 1College of Animal Science and Technology, Northwest A&F University, Yangling, Shaanxi, China.

Methods in Molecular Biology (Clifton, N.J.)
|August 30, 2018
PubMed
Summary

This study introduces a novel suicidal zinc-finger nuclease (ZFN) system with a surrogate reporter, significantly reducing cellular toxicity by ~40% while enabling efficient enrichment of genetically modified cells.

Keywords:
Cellular toxicityDouble-stranded breaksHomologous recombination repairOff-targetZinc-finger nucleases

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Area of Science:

  • Molecular Biology
  • Biotechnology
  • Genetics Engineering

Background:

  • Engineered nucleases like zinc-finger nucleases (ZFNs) and TALE nucleases (TALENs) are powerful genome editing tools.
  • However, their application is limited by high cellular toxicity and off-targeting effects.
  • Enriching genetically modified cells is challenging due to the lack of effective selection markers.

Purpose of the Study:

  • To develop a genome editing system with reduced cellular toxicity and improved selection of modified cells.
  • To integrate a surrogate reporter enrichment strategy into a suicidal ZFN system.
  • To evaluate the efficacy and safety of the novel system.

Main Methods:

  • A suicidal ZFN expression system was designed with ZFN expression cassettes flanked by target sites.
  • A surrogate reporter (EGFP) was incorporated for cell enrichment.
  • Cellular toxicity and ZFN activity were compared between the modified and original systems.

Main Results:

  • The modified suicidal ZFN system demonstrated comparable ZFN activity to the original system.
  • A significant reduction in cellular toxicity (~40%) was observed with the new system.
  • The surrogate reporter facilitated efficient enrichment of genetically modified cells via EGFP analysis.

Conclusions:

  • The novel suicidal ZFN expression system coupled with a surrogate reporter effectively reduces cellular toxicity in genome editing.
  • This approach simplifies the enrichment of genetically modified cells, enhancing the utility of ZFN technology.
  • The findings offer a promising strategy for safer and more efficient genome editing applications.