CRISPR-U™ Gene Knockout Cell Line Strategy

ACE2 Gene Knockout HL-60 Cell 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 ACE2 Knockout model in HL-60 cell line by CRISPR-U™-mediated genome engineering.
Target gene info
Official symbol ACE2
Gene id 59272
Organism Homo sapiens
Official full symbol angiotensin I converting enzyme 2
Gene type protein-coding
Also known as ACEH
Summary The protein encoded by this gene belongs to the angiotensin-converting enzyme family of dipeptidyl carboxydipeptidases and has considerable homology to human angiotensin 1 converting enzyme. This secreted protein catalyzes the cleavage of angiotensin I into angiotensin 1-9, and angiotensin II into the vasodilator angiotensin 1-7. ACE2 is known to be expressed in various human organs, and its organ- and cell-specific expression suggests that it may play a role in the regulation of cardiovascular and renal function, as well as fertility. In addition, the encoded protein is a functional receptor for the spike glycoprotein of the human coronavirus HCoV-NL63 and the human severe acute respiratory syndrome coronaviruses, SARS-CoV and SARS-CoV-2, the causative agent of coronavirus disease-2019 (COVID-19).
Genomic regions Chromosome X
Cell line info
Cell name HL-60
Tissue peripheral blood
Organism Human
Morphology myeloblastic
Culture properties suspension
Disease acute promyelocytic leukemia
Derivation HL-60 is a promyelocytic cell line derived by S.J. Collins, et al. Peripheral blood leukocytes were obtained by leukopheresis from a 36-year-old Caucasian female with acute promyelocytic leukemia.
Complete growth medium Complete Growth MediumThe base medium for this cell line is ATCC-formulated Iscove's Modified Dulbecco's Medium, Catalog No. 30-2005.  To make the complete growth medium, add the following components to the base medium:  ·fetal bovine serum to a final concentration of 20%This medium is formulated for use with a 5% CO2 in air atmosphere.ATCC tested fetal bovine serum is available as ATCC Catalog No. 30-2020.
Subcultivation ratio Subcultivation Ratio:Cultures can be maintained by the addition of fresh medium or replacement of medium. Alternatively, cultures can be established by centrifugation with subsequent resuspension at 1 X 105 viable cells/mL. Do not allow cell concentration to exceed 1 X 106 cells/mL.Corning® T-75 flasks (catalog #431464) are recommended for subculturing this product. Interval: Maintain cell density between 1 x 105and 1 x 106 viable cells/mL. Medium Renewal: Every 2 to 3 days
Strategy Summary
This gene has 8 protein coding transcripts:
Name Transcript ID bp Protein Biotype CCDS UniProt Match RefSeq Match Flags
ACE2-202 ENST00000427411.2 7587 805aa Protein coding CCDS14169 Q9BYF1-1 - TSL:1, GENCODE basic, APPRIS P2,
ACE2-207 ENST00000678073.1 7487 805aa Protein coding CCDS14169 - - GENCODE basic, APPRIS P2,
ACE2-211 ENST00000680121.1 7413 805aa Protein coding CCDS14169 - - GENCODE basic, APPRIS P2,
ACE2-201 ENST00000252519.8 3339 805aa Protein coding CCDS14169 Q9BYF1-1 NM_001371415.1 TSL:1, GENCODE basic, APPRIS P2, MANE Select v0.92,
ACE2-205 ENST00000677282.1 6372 459aa Protein coding - - - GENCODE basic,
ACE2-210 ENST00000679278.1 3149 786aa Protein coding - - - GENCODE basic, APPRIS ALT2,
ACE2-206 ENST00000678046.1 2723 771aa Protein coding - - - GENCODE basic, APPRIS ALT2,
ACE2-209 ENST00000679212.1 2642 772aa Protein coding - - - GENCODE basic, APPRIS ALT2,
ACE2-208 ENST00000679162.1 2730 786aa Nonsense mediated decay - - - -
ACE2-203 ENST00000471548.5 998 112aa Nonsense mediated decay - - - CDS 5' incomplete, TSL:2,
ACE2-204 ENST00000473851.1 786 No protein Retained intron - - - TSL:3,
Ubigene Red Cotton Transcript
Strategy Click to get
Red Cotton™ Assessment    
Project Difficulty Level unknown
Cell Line HL-60
Target Gene ACE2
Colony Formation Intermediate
This KO Strategy loading
Red Cotton™ Notes Ubigene has successfully modified HL-60 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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