CRISPR-U™ Gene Knockout Cell Line
Strategy
ACE2 Gene Knockout C2C12 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 C2C12 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 | C2C12 |
| Tissue | muscle |
| Organism | Mouse |
| Morphology | myoblast |
| Culture properties | adherent |
| Derivation | This is a subclone (produced by H. Blau, et al) of the mouse myoblast cell line established by D. Yaffe and O. Saxel. |
| Complete growth medium | Complete growth medium described above supplemented with 5% (v/v) DMSO. Cell culture tested DMSO is available as ATCC Catalog No. 4-X. |
| Subcultivation ratio | Subcultivation Ratio:IMPORTANT - DO NOT ALLOW CULTURES TO BECOME CONFLUENT. Cultures must not be allowed to become confluent as this will deplete the myoblastic population in the culture. Myotube formation is enhanced when the medium is supplemented with 10% horse serum instead of fetal bovine serum. Remove and discard culture medium. Briefly rinse the cell layer with 0.25% (w/v) Trypsin- 0.53 mM EDTA solution to remove all traces of serum which contains trypsin inhibitor. Add 2.0 to 3.0 mL of Trypsin-EDTA solution to flask and observe cells under an inverted microscope until cell layer is dispersed (usually within 5 to 15 minutes). Note: To avoid clumping do not agitate the cells by hitting or shaking the flask while waiting for the cells to detach. Cells that are difficult to detach may be placed at 37°C to facilitate dispersal. Add 6.0 to 8.0 mL of complete growth medium and aspirate cells by gently pipetting. Add appropriate aliquots of the cell suspension to new culture vessels. Inoculate at a cell concentration between 1.5 X 10 5 and 1.0 X 10 6 viable cells/75 cm2.Corning® T-75 flasks (catalog #430641) are recommended for subculturing this product. Incubate cultures at 37°C. Medium Renewal: Every two to three 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 unknown
| Cell Line | C2C12 |
| Target Gene | ACE2 |
| Colony Formation | Simple |
| This KO Strategy | loading |
| Red Cotton™ Notes | Ubigene has successfully modified C2C12 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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