CRISPR-U™ Gene Knockout Cell Line Strategy

RAF1 Gene Knockout Strategy

CRISPR-U™ technology (CRISPR based), developed by Ubigene, is more efficient than general CRISPR/Cas9 technology in double-strand breaking and homologous recombination. With CRISPR-U™, Ubigene has successfully edited over 3000 genes on more than 100 types of cell lines.
Objective
To create a Human RAF1 Knockout model in cell line by CRISPR-U™-mediated genome engineering.
Target gene info
Official symbol RAF1
Gene id 5894
Organism Homo sapiens
Official full symbol Raf-1 proto-oncogene, serine/threonine kinase
Gene type protein-coding
Also known as CMD1NN, CRAF, NS5, Raf-1, c-Raf
Summary This gene is the cellular homolog of viral raf gene (v-raf). The encoded protein is a MAP kinase kinase kinase (MAP3K), which functions downstream of the Ras family of membrane associated GTPases to which it binds directly. Once activated, the cellular RAF1 protein can phosphorylate to activate the dual specificity protein kinases MEK1 and MEK2, which in turn phosphorylate to activate the serine/threonine specific protein kinases, ERK1 and ERK2. Activated ERKs are pleiotropic effectors of cell physiology and play an important role in the control of gene expression involved in the cell division cycle, apoptosis, cell differentiation and cell migration. Mutations in this gene are associated with Noonan syndrome 5 and LEOPARD syndrome 2.
Genomic regions Chromosome 3
Strategy Summary
This gene has 3 protein coding transcripts:
Name Transcript ID bp Protein Biotype CCDS UniProt Match RefSeq Match Flags
RAF1-205 ENST00000442415.7 3251 668aa Protein coding CCDS87047 P04049-2 NM_001354689.3 TSL:5, GENCODE basic, APPRIS ALT1, MANE Select v0.92,
RAF1-201 ENST00000251849.9 3191 648aa Protein coding CCDS2612 P04049-1 - TSL:1, GENCODE basic, APPRIS P4,
RAF1-204 ENST00000432427.2 1831 527aa Protein coding - H7C155 - CDS 5' incomplete, TSL:5,
RAF1-203 ENST00000423275.5 2354 69aa Nonsense mediated decay - A0A0B4J1W9 - TSL:2,
RAF1-202 ENST00000416093.1 962 69aa Nonsense mediated decay - A0A0B4J1W9 - TSL:2,
RAF1-212 ENST00000494557.1 3205 No protein Retained intron - - - TSL:1,
RAF1-207 ENST00000465826.5 1518 No protein Retained intron - - - TSL:3,
RAF1-208 ENST00000471449.1 804 No protein Retained intron - - - TSL:2,
RAF1-211 ENST00000492690.1 633 No protein Retained intron - - - TSL:3,
RAF1-209 ENST00000475353.1 599 No protein Retained intron - - - TSL:4,
RAF1-210 ENST00000491290.1 572 No protein Retained intron - - - TSL:4,
RAF1-206 ENST00000460610.1 484 No protein Retained intron - - - TSL:3,
Ubigene Red Cotton Transcript
Strategy
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Red Cotton™ Assessment    
Project Difficulty Level unknown
Target Gene RAF1
This KO Strategy loading
Red Cotton™ Notes Gene RAF1 had been KO in hct116 cell line.
Aforementioned information comes from Ubigene database. Different origin of cell lines may have different condition. Ubigene reserved all the right for final explanation.
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Work flow
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