CRISPR-U™ Gene Knockout Cell Line
Strategy
ACE2 Gene Knockout KYSE-150 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 KYSE-150 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 | KYSE-150 |
| Tissue | Esophagus |
| Organism | Human |
| Morphology | Epithelial |
| Culture properties | Monolayer, adherent |
| Disease | esophageal squamous cell carcinoma |
| Derivation | upper (cervical) esophagus |
| Complete growth medium | 90%RPMI-1640+10%FBS |
| Subcultivation ratio | Subculturing: Remove and discard culture medium. Briefly rinse the cell layer with DPBS solution to remove all traces of serum that contains trypsin inhibitor. Add 1.0 to 2.0 mL of Trypsin-EDTA solution to flask and observe cells under an inverted microscope until cell layer is dispersed (usually within 2 to 3 minutes). Cells that are difficult to detach may be placed at 37°C to facilitate dispersal. Add 4.0 to 6.0 mL of complete growth medium and aspirate cells by gently pipetting. Add appropriate aliquots of the cell suspension to new culture vessels. Ratio: 1:2-1:5. 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, |
Strategy
Click to get
Red Cotton™ Assessment
Project Difficulty Level 1
| Cell Line | KYSE-150 |
| Target Gene | ACE2 |
| Colony Formation | Simple |
| AVG. Gene Copy Number | ≈3.0 |
| Transfection Rate | ≥70.0% |
| This KO Strategy | loading |
| Red Cotton™ Notes | Ubigene has successfully modified KYSE-150 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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