Experimenting on human zygotes using clustered regularly interspaced short palindromic repeat genome editing technology

from For the sake of reproducibility - a technical replicate by Tumor Necrosis Factor-alpha

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Clustered regularly interspaced short palindromic repeat associated system
An endonuclease complex identified as a part of the immune system of streptococcus pyogenes
A powerful tool in the context of genome editing, gene therapy and gene functional investigation
Yet which still needs to be fully understood in order to minimise unintended mutation and non-specific target recognition

Reprogramming of guiding gRNA sequences
Targeting of a mutated gene of interest
A precise DNA repair mechanism
Potential clinical applications for the CRISPR/Cas9 system

Cultured cells from tripronuclear human zygotes were (ab)used to further investigate the system
Low efficiency in homologous (recombination) directed repair and off-target cleavage observation
Endogenous homologous gene competing with the exogenous donour, causing untoward mutations
Improving the specificity and accuracy of the genome editing method may necessitate years of additional experimentation

Reprogramming of guiding gRNA sequences
Off-target effects, mutations and mosaicism
A not-so precise genome editing mechanism
Potential caveats for the CRISPR/Cas9 system

An otherwise low efficiency of homologous recombination directed repair, due to competing endogenous paralogs and pseudogenes
Combined with off-target cleavage and interfering repair template, mosaic tripronuclear zygotes
hindered by untoward mutations

Ref : Puping L et al. “CRISPR/Cas9-mediated gene editing in human tripronuclear zygotes.” Protein & Cell 6(5): 363–372. (2015)

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from For the sake of reproducibility - a technical replicate, released January 24, 2018

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Scientists, academics, gorefiends and grindheads of the world rejoice: Tumor Necrosis Factor-alpha unleashes its take on full blown Carcass-worship, topped with unique lyrical themes inspired by the most recent knowledge in state-of-the-art scientific research from the fields of cellular and molecular biology. ... more

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