CRISPR-U™ Gene Knockout Cell Line Strategy

AKT1 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 AKT1 Knockout model in cell line by CRISPR-U™-mediated genome engineering.
Target gene info
Official symbol AKT1
Gene id 207
Organism Homo sapiens
Official full symbol AKT serine/threonine kinase 1
Gene type protein-coding
Also known as AKT, PKB, PKB-ALPHA, PRKBA, RAC, RAC-ALPHA
Summary This gene encodes one of the three members of the human AKT serine-threonine protein kinase family which are often referred to as protein kinase B alpha, beta, and gamma. These highly similar AKT proteins all have an N-terminal pleckstrin homology domain, a serine/threonine-specific kinase domain and a C-terminal regulatory domain. These proteins are phosphorylated by phosphoinositide 3-kinase (PI3K). AKT/PI3K forms a key component of many signalling pathways that involve the binding of membrane-bound ligands such as receptor tyrosine kinases, G-protein coupled receptors, and integrin-linked kinase. These AKT proteins therefore regulate a wide variety of cellular functions including cell proliferation, survival, metabolism, and angiogenesis in both normal and malignant cells. AKT proteins are recruited to the cell membrane by phosphatidylinositol 3,4,5-trisphosphate (PIP3) after phosphorylation of phosphatidylinositol 4,5-bisphosphate (PIP2) by PI3K. Subsequent phosphorylation of both threonine residue 308 and serine residue 473 is required for full activation of the AKT1 protein encoded by this gene. Phosphorylation of additional residues also occurs, for example, in response to insulin growth factor-1 and epidermal growth factor. Protein phosphatases act as negative regulators of AKT proteins by dephosphorylating AKT or PIP3. The PI3K/AKT signalling pathway is crucial for tumor cell survival. Survival factors can suppress apoptosis in a transcription-independent manner by activating AKT1 which then phosphorylates and inactivates components of the apoptotic machinery. AKT proteins also participate in the mammalian target of rapamycin (mTOR) signalling pathway which controls the assembly of the eukaryotic translation initiation factor 4F (eIF4E) complex and this pathway, in addition to responding to extracellular signals from growth factors and cytokines, is disregulated in many cancers. Mutations in this gene are associated with multiple types of cancer and excessive tissue growth including Proteus syndrome and Cowden syndrome 6, and breast, colorectal, and ovarian cancers. Multiple alternatively spliced transcript variants have been found for this gene.
Genomic regions Chromosome 14
Strategy Summary
This gene has 9 protein coding transcripts:
Name Transcript ID bp Protein Biotype CCDS UniProt Match RefSeq Match Flags
AKT1-208 ENST00000554581.5 3916 480aa Protein coding CCDS9994 P31749-1 - TSL:1, GENCODE basic, APPRIS P1,
AKT1-202 ENST00000402615.6 3913 480aa Protein coding CCDS9994 P31749-1 - TSL:1, GENCODE basic, APPRIS P1,
AKT1-214 ENST00000555528.5 3066 480aa Protein coding CCDS9994 P31749-1 - TSL:1, GENCODE basic, APPRIS P1,
AKT1-219 ENST00000649815.2 2957 480aa Protein coding CCDS9994 P31749-1 NM_001382430.1 GENCODE basic, APPRIS P1, MANE Select v0.92,
AKT1-201 ENST00000349310.7 2866 480aa Protein coding CCDS9994 P31749-1 - TSL:1, GENCODE basic, APPRIS P1,
AKT1-203 ENST00000407796.7 2779 480aa Protein coding CCDS9994 P31749-1 - TSL:1, GENCODE basic, APPRIS P1,
AKT1-211 ENST00000554848.5 1595 480aa Protein coding CCDS9994 P31749-1 - TSL:1, GENCODE basic, APPRIS P1,
AKT1-207 ENST00000554192.5 922 184aa Protein coding - G3V2I6 - CDS 5' incomplete, TSL:2,
AKT1-213 ENST00000555458.5 705 170aa Protein coding - G3V3X1 - CDS 5' incomplete, TSL:2,
AKT1-209 ENST00000554585.5 858 146aa Nonsense mediated decay - A0A087WY56 - CDS 5' incomplete, TSL:3,
AKT1-204 ENST00000544168.5 1564 No protein Processed transcript - - - TSL:2,
AKT1-206 ENST00000553797.1 783 No protein Processed transcript - - - TSL:3,
AKT1-212 ENST00000555380.1 534 No protein Processed transcript - - - TSL:4,
AKT1-216 ENST00000557494.1 408 No protein Processed transcript - - - TSL:2,
AKT1-217 ENST00000557552.1 8396 No protein Retained intron - - - TSL:5,
AKT1-218 ENST00000610370.1 3187 No protein Retained intron - - - TSL:2,
AKT1-205 ENST00000553506.5 2830 No protein Retained intron - - - TSL:2,
AKT1-215 ENST00000556836.1 592 No protein Retained intron - - - TSL:4,
AKT1-210 ENST00000554826.1 572 No protein Retained intron - - - TSL:3,
Ubigene Red Cotton Transcript
Strategy
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Red Cotton™ Assessment    
Project Difficulty Level unknown
Target Gene AKT1
This KO Strategy loading
Red Cotton™ Notes Gene AKT1 had been KO in hela 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.
Special deals for this gene:

$49

Single gRNA plasmid off-shelf

$599

Single gRNA lentivirus

Work flow
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