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zebrafish cpf1

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https://read.qxmd.com/read/30886848/expanding-the-crispr-toolbox-in-zebrafish-for-studying-development-and-disease
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Kaili Liu, Cassidy Petree, Teresa Requena, Pratishtha Varshney, Gaurav K Varshney
The study of model organisms has revolutionized our understanding of the mechanisms underlying normal development, adult homeostasis, and human disease. Much of what we know about gene function in model organisms (and its application to humans) has come from gene knockouts: the ability to show analogous phenotypes upon gene inactivation in animal models. The zebrafish ( Danio rerio ) has become a popular model organism for many reasons, including the fact that it is amenable to various forms of genetic manipulation...
2019: Frontiers in Cell and Developmental Biology
https://read.qxmd.com/read/29964176/optimized-crispr-cpf1-system-for-genome-editing-in-zebrafish
#2
Juan P Fernandez, Charles E Vejnar, Antonio J Giraldez, Romain Rouet, Miguel A Moreno-Mateos
The impact of the CRISPR-Cas biotechnological systems has recently broadened the genome editing toolbox available to different model organisms further with the addition of new efficient RNA-guided endonucleases. We have recently optimized CRISPR-Cpf1 (renamed Cas12a) system in zebrafish. We showed that (i) in the absence of Cpf1 protein, crRNAs are unstable and degraded in vivo, and CRISPR-Cpf1 RNP complexes efficiently mutagenize the zebrafish genome; and (ii) temperature modulates Cpf1 activity especially affecting AsCpf1, which experiences a reduced performance below 37 °C...
November 1, 2018: Methods: a Companion to Methods in Enzymology
https://read.qxmd.com/read/29222508/crispr-cpf1-mediates-efficient-homology-directed-repair-and-temperature-controlled-genome-editing
#3
Miguel A Moreno-Mateos, Juan P Fernandez, Romain Rouet, Charles E Vejnar, Maura A Lane, Emily Mis, Mustafa K Khokha, Jennifer A Doudna, Antonio J Giraldez
Cpf1 is a novel class of CRISPR-Cas DNA endonucleases, with a wide range of activity across different eukaryotic systems. Yet, the underlying determinants of this variability are poorly understood. Here, we demonstrate that LbCpf1, but not AsCpf1, ribonucleoprotein complexes allow efficient mutagenesis in zebrafish and Xenopus. We show that temperature modulates Cpf1 activity by controlling its ability to access genomic DNA. This effect is stronger on AsCpf1, explaining its lower efficiency in ectothermic organisms...
December 8, 2017: Nature Communications
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