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Construction of a GLI3 compound heterozygous knockout human embryonic stem cell line WAe001-A-20 by CRISPR/Cas9 editing.

Stem cell research | 2018

The human GLI3 protein has a dual function as a transcriptional activator or repressor of hedgehog signaling, depending on the proteolytic processing forms of GLI3. In this study, we established a compound heterozygous GLI3 mutant human embryonic stem cell line (WAe001-A-20) through CRISPR/Cas9 editing. The WAe001-A-20 cells carried two deletions on two different alleles of exon 2 of GLI3, respectively, which resulted in a frame shift and early termination in the translation of GLI3. Moreover, WAe001-A-20 maintains a normal karyotype, parental cell morphology, pluripotent phenotype and the ability to differentiate into three germ layers. Resource table.

Pubmed ID: 30278376 RIS Download

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This is a list of tools and resources that we have found mentioned in this publication.


Addgene (tool)

RRID:SCR_002037

Non-profit plasmid repository dedicated to helping scientists around the world share high-quality plasmids. Facilitates archiving and distributing DNA-based research reagents and associated data to scientists worldwide. Repository contains over 65,000 plasmids, including special collections on CRISPR, fluorescent proteins, and ready-to-use viral preparations. There is no cost for scientists to deposit plasmids, which saves time and money associated with shipping plasmids themselves. All plasmids are fully sequenced for validation and sequencing data is openly available. We handle the appropriate Material Transfer Agreements (MTA) with institutions, facilitating open exchange and offering intellectual property and liability protection for depositing scientists. Furthermore, we curate free educational resources for the scientific community including a blog, eBooks, video protocols, and detailed molecular biology resources.

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PeproTech (tool)

RRID:SCR_006802

An Antibody supplier

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Open Reading Frame Finder (tool)

RRID:SCR_016643

Software tool to search for open reading frames (ORFs) in the DNA sequence. The program returns the range of each ORF, along with its protein translation. Used to search newly sequenced DNA for potential protein encoding segments, verify predicted protein. Limited to the subrange of the query sequence up to 50 kb long.

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Anti-Gli3 antibody (antibody)

RRID:AB_2732891

This monoclonal targets Gli3

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Human PDX-1/IPF1 Affinity Purified Polyclonal Ab (antibody)

RRID:AB_355257

This polyclonal targets Human PDX-1/IPF1 Affinity Purified Ab

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Alpha Tubulin antibody (antibody)

RRID:AB_2687491

This monoclonal targets Alpha Tubulin

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GAPDH antibody (antibody)

RRID:AB_2631281

This unknown targets

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SSEA-4 (813-70) (antibody)

RRID:AB_628289

This monoclonal targets Human Unknown

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TRA-1-60 (TRA-1-60) (antibody)

RRID:AB_628385

This monoclonal targets Human PODXL

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Oct3/4 Antibody (C-10) (antibody)

RRID:AB_628051

This monoclonal targets Oct-3/4

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Anti-Gli3 antibody (antibody)

RRID:AB_2732891

This monoclonal targets Gli3

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