genome wide crispr cas9 knockout screen human crispr knockout pooled brunello library  (Addgene inc)

Journal: Journal of Translational Medicine

Article Title: CRISPR-Cas and CRISPR-based screening system for precise gene editing and targeted cancer therapy

doi: 10.1186/s12967-024-05235-2

Figure Lengend Snippet: Schematic representative of CRISPR/Cas loci in Class 1 and Class 2 system. Class 1 system show multi-component effectors, while the Class 2 system have one effector. Three subgroups of Class 2 CRISPR systems are presented. Representative Type II-A CRISPR protein contains: Streptococcus pyogenes Cas9 (SpCas9), Staphylococcus aureus Cas9 (SaCas9) and Streptococcus thermophilus Cas9 (StrCas9), all of which have the tracrRNA sequences. Type V CRISPR, which comprises Cas12a, Cas12b and Cas12c, exhibits distinct genome structures. Cas12b has the tracrRNA structure, while Cas12c only has one assistant protein cas1 for genome editing. Cas14 subgroup is not depicted in this figure. Type VI CRISPR systems show few assistant proteins to identify RNA virus, however, type VI-B has csx27 and csx28 proteins to regulate nuclease activity. Illustrated according to Ref [ , , , ].

Article Snippet: Cas9 , Addgene #73,179 , CRC cell line SW480 CRC organoids , CRISPRko , Drug resistance , [ ] .

Techniques: CRISPR, Virus, Activity Assay

Journal: Journal of Translational Medicine

Article Title: CRISPR-Cas and CRISPR-based screening system for precise gene editing and targeted cancer therapy

doi: 10.1186/s12967-024-05235-2

Figure Lengend Snippet: Summary of Cas9 proteins and modified nCas9 and dCas9 genome editing tools. (A) PAM for SpCas9 is NGG, while PAM for SaCas9 is NNGRRT with the ability to cut DNA double helix. (B) Mutation of D10A leads to the formation of nCas9 while both mutations generate dCas9 protein. (C) nCas9 can be applied for base editing such as CBE and ABE, also for Base editor and developed as NICER to repair heterogenous mutation. (D) dCas9 was modified to generate CRISPRi, CRISPRa and CRISPR labeling tools. dCas9: dead Cas9. nCas9: nickase Cas9. CBE: Cytosine Base Editor, ABE: Adenine Base Editor. RT: reverse transcriptase. pegRNA: prime editing guide RNA.

Article Snippet: Cas9 , Addgene #73,179 , CRC cell line SW480 CRC organoids , CRISPRko , Drug resistance , [ ] .

Techniques: Modification, Mutagenesis, CRISPR, Labeling, Reverse Transcription

Journal: Journal of Translational Medicine

Article Title: CRISPR-Cas and CRISPR-based screening system for precise gene editing and targeted cancer therapy

doi: 10.1186/s12967-024-05235-2

Figure Lengend Snippet: CRISPR screening ex vivo for cancer research

Article Snippet: Cas9 , Addgene #73,179 , CRC cell line SW480 CRC organoids , CRISPRko , Drug resistance , [ ] .

Techniques: CRISPR, Ex Vivo, Biomarker Discovery, Ubiquitin Proteomics, Activation Assay, Plasmid Preparation, Drug discovery

Journal: Journal of Translational Medicine

Article Title: CRISPR-Cas and CRISPR-based screening system for precise gene editing and targeted cancer therapy

doi: 10.1186/s12967-024-05235-2

Figure Lengend Snippet: CRISPR screening in vivo for cancer therapy

Article Snippet: Cas9 , Addgene #73,179 , CRC cell line SW480 CRC organoids , CRISPRko , Drug resistance , [ ] .

Techniques: CRISPR, In Vivo, Multiplex Assay, Genome Wide