An efficient method for generation of Knock-out human embryonic stem cells using CRISPR/Cas9 system (Record no. 50857)

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fixed length control field 02138nam a2200253Ia 4500
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control field MX-MdCICY
005 - DATE AND TIME OF LATEST TRANSACTION
control field 20250625160156.0
040 ## - CATALOGING SOURCE
Transcribing agency CICY
090 ## - LOCALLY ASSIGNED LC-TYPE CALL NUMBER (OCLC); LOCAL CALL NUMBER (RLIN)
Classification number (OCLC) (R) ; Classification number, CALL (RLIN) (NR) B-16687
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245 10 - TITLE STATEMENT
Title An efficient method for generation of Knock-out human embryonic stem cells using CRISPR/Cas9 system
490 0# - SERIES STATEMENT
Volume/sequential designation Stem Cells Dev., doi: 10.1089/scd.2017.0058., 2017
520 3# - SUMMARY, ETC.
Summary, etc. Human embryonic stem cells (hESCs)represent promising tool to study functions of genes during development, to model diseases, and to even develop therapies when combined with gene editing techniques such as CRISPR/Cas9 system. However, the process of disruption of gene expression by generation of null alleles is often inefficient and tedious. To circumvent these limitations, we developed a simple and efficient protocol to permanently downregulate expression of gene of interest in hESCs using CRISPR/Cas9. We selected p53 for our proof of concept experiments. The methodology is based on series of hESC transfection, which leads to efficient downregulation of p53 expression even in polyclonal population (p53 Low cells), here proven by a loss of regulation of the expression of p53 target gene, microRNA miR-34a. We demonstrate that our approach achieves over 80 percent efficiency in generating hESC clonal sublines that do not express p53 protein. Importantly, we document by a set of functional experiments that such genetically modified hESC do retain typical stem cells characteristics. In summary, we provide a simple and robust protocol to efficiently target expression of gene of interest in hESCs that can be useful for laboratories aiming to employ gene editing in their hESC applications/protocols.
650 14 - SUBJECT ADDED ENTRY--TOPICAL TERM
Topical term or geographic name entry element STEM CELLS (HESCS)
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Personal name Bohaciakova D
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Personal name Renzova T
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Personal name Fedorova V
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Personal name Barak M
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Personal name Kunova Bosakova M
700 12 - ADDED ENTRY--PERSONAL NAME
Personal name Hampl A
700 12 - ADDED ENTRY--PERSONAL NAME
Personal name Cajanek L
856 40 - ELECTRONIC LOCATION AND ACCESS
Uniform Resource Identifier <a href="https://drive.google.com/file/d/1wPxY7OGND9Lae04f8fgvGFqsnOxD-LDK/view?usp=drivesdk">https://drive.google.com/file/d/1wPxY7OGND9Lae04f8fgvGFqsnOxD-LDK/view?usp=drivesdk</a>
Public note Para ver el documento ingresa a Google con tu cuenta: @cicy.edu.mx
942 ## - ADDED ENTRY ELEMENTS (KOHA)
Source of classification or shelving scheme Clasificación local
Koha item type Documentos solicitados
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  Clasificación local     Ref1 CICY CICY Documento préstamo interbibliotecario 25.06.2025   B-16687 25.06.2025 25.06.2025 Documentos solicitados