PXD042667

PXD042667 is an original dataset announced via ProteomeXchange.

Title	Using a Dual CRISPR/Cas9 Approach to Gain Insight into the Role of LRP1B in Glioblastoma
Description	LRP1B remains one of the most altered genes in cancer, although its relevance in cancer biology is still unclear. Recent advances in gene editing techniques, particularly CRISPR/Cas9 systems, offer new opportunities to evaluate the function of large genes, like LRP1B. Using a dual sgRNA CRISPR/Cas9 gene editing approach, this study aimed to assess the impact of disrupting LRP1B in glioblastoma cell biology. Four sgRNAs were designed for the dual targeting of two LRP1B exons (1 and 85). The U87 glioblastoma (GB) cell line was transfected with CRISPR/cas9 PX459 vectors. To assess LRP1B gene alterations and expression, PCR, Sanger DNA sequencing, qRT-PCR were carried out. Three clones (clones B9, E6 and H7) were further evaluated. All clones presented altered cellular morphology, increased cellular and nuclear size and changes in ploidy. Two clones (E6 and H7) showed a significant decrease in cell growth, both in vitro and in the in vivo CAM assay. Proteomic analysis of the clones’ secretome identified differentially ex-pressed proteins, that had not been previously associated with LRP1B alterations. This study demonstrates that the dual sgRNA CRISPR/Cas9 strategy can effectively edit LRP1B in GB cells, providing new insights into the impact of LRP1B deletions in GBM biology.
HostingRepository	PRIDE
AnnounceDate	2023-11-14
AnnouncementXML	Submission_2023-11-14_09:11:41.578.xml
DigitalObjectIdentifier
ReviewLevel	Peer-reviewed dataset
DatasetOrigin	Original dataset
RepositorySupport	Unsupported dataset by repository
PrimarySubmitter	Hugo Osorio
SpeciesList	scientific name: Homo sapiens (Human); NCBI TaxID: 9606;
ModificationList	monohydroxylated residue; iodoacetamide derivatized residue
Instrument	Q Exactive

Revision	Datetime	Status	ChangeLog Entry
0	2023-06-02 07:30:36	ID requested
1	2023-10-24 13:22:41	announced
⏵ 2	2023-11-14 09:11:44	announced	2023-11-14: Updated project metadata.

Peixoto J, Pr, í, ncipe C, Pestana A, Os, ó, rio H, Pinto MT, Prazeres H, Soares P, Lima RT, Using a Dual CRISPR/Cas9 Approach to Gain Insight into the Role of LRP1B in Glioblastoma. Int J Mol Sci, 24(14):(2023) [pubmed]

submitter keyword: glioblastoma, ploidy, CRISPR/Cas9,LRP1B , secretome

Raquel Lima
contact affiliation	i3S—Instituto de Investigação e Inovação em Saúde, Universidade do Porto, 4200-135 Porto, Portugal
contact email	rlima@ipatimup.pt
lab head
Hugo Osorio
contact affiliation	i3S - Institute for Research and Innovation in Health, University of Porto
contact email	hosorio@i3s.up.pt
dataset submitter

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PRIDE project URI

• PRIDE
  1. PXD042667
     1. Label: PRIDE project
     2. Name: Using a Dual CRISPR/Cas9 Approach to Gain Insight into the Role of LRP1B in Glioblastoma