LIST OF PARTS
Basic Parts
| Name | Type | Description | Designers | Information |
|---|---|---|---|---|
| BBa_K4501000 | RNA | anti-myc shRNA 1741-1761 | Pau | shRNA against myc to silence the gene expression. |
| BBa_K4501001 | RNA | anti-myc shRNA 1741-1761 C/D-box | Pau | shRNA against myc with a C/D-box sequence on the 3’ terminus. |
| BBa_K4501002 | RNA | anti-myc shRNA 1741-1761 in-loop C/D-box | Pau | shRNA against myc with a C/D-box sequence in substitution to the traditional loop. |
| BBa_K4501003 | Other | AAVS1 5’ homology arm | Eva | Contains the homology sequence for AAVS1 site to achieve safe integration. |
| BBa_K4501004 | Other | AAVS1 3’ homology arm | Eva | Contains the homology sequence for AAVS1 site to achieve safe integration. |
| BBa_K4501005 | Device | MSCV 5’ module | Pau | 5’ LTR and other elements to help with the integration of the transgene. |
| BBa_K4501006 | Device | MSCV 3’ module | Pau | 3’ LTR and other elements to help with the integration of the transgene. |
| BBa_K4501007 | Reporter | HygR | Pau | Gene providing hygromycin resistance, used for selection. |
| BBa_K4501008 | Other | cPPT/CTS | Pau | Increases transfection and transgene expression. |
| BBa_K4501009 | Cell | CD19-L | Pau | Ligand of CD19 surface marker, used to target B cells. |
| Bba_K4501010 | Cell | Connexin 43 S368A | Pau | Aquaporin channel used to increase the flow of shRNA to exosomes. |
Our favourite Basic Part: anti-myc shRNA in-loop C/D-box (BBa_K4501002)
Amongst our basic parts BBa_K4501002 is our favourite. With this part, we believe that we have found a stable, versatile and elegant system to label shRNAs with the C/D-box motif. By substituting the traditionally used loop for the C/D-box motif, labelling of shRNAs is achieved in a convenient and advantageous fashion. Furthermore, the substitution of the traditional loop with the C/D-box loop motif even increases the stability of the shRNA double-stranded monomer, a positive feature for the shRNA function.
Composite Parts
| Name | Type | Description | Designers | Information |
|---|---|---|---|---|
| Bba_K4501010 | Device | rgasra | Pau | Contains exosome booster genes such as STEAP3, SDC4 and NadB and a puromycin resistance gene for selection. |
| BBa_K4501012 | Composite | Exosome booster 2 | Eva | Contains exosome booster gene nSMase and a hygromycin resistance gene for selection. |
| BBa_K4501013 | Composite | CD63_His-tag | Jaume | Contains the tetraspannin CD63 labelled with 6x His-tag coupled to a RFP reporter. |
| BBa_K4501014 | Composite | CD63_His-tag L7Ae (1) | Jaume | Contains the tetraspannin CD63 labelled with 6x His-tag and a L7Ae unit for RNA laoding, coupled to a RFP reporter. |
| BBa_K4501015 | Composite | CD63_His-tag L7Ae (2) | Jaume | Contains the tetraspannin CD63 labelled with 6x His-tag and two L7Ae units for RNA laoding, coupled to a RFP reporter. |
| BBa_K4501016 | Composite | CD63_His-tag L7Ae (3) | Jaume | Contains the tetraspannin CD63 labelled with 6x His-tag and two L7Ae units for RNA laoding, coupled to a RFP reporter. | BBa_K4501017 | Composite | Lamp2B-CD19-L | Pau | Contains the fusion protein Lamp2b-CD19-L to provide B cell tropism to our exosomes, coupled to a BFP reporter. | BBa_K4501018 | Composite | myc shRNA | Pau | Contains the expression cassette to express the shRNA and a GFP reporter. | BBa_K4501019 | Composite | myc shRNA C/D-box | Pau | Contains the expression cassette to express the shRNA and a GFP reporter. | BBa_K4501020 | Composite | myc shRNA in-loop C/D-box | Pau | Contains the expression cassette to express the shRNA and a GFP reporter. | BBa_K4501021 | Composite | Cytosolic delivery helper | Pau | Contains the mutant S384A version of Cx43 | BBa_K4501022 | Backbone | pJUMP lentiviral transfer plasmid | Pau | Improvement of the pJUMP29-1A KanR L1 backbone with both 5’ and 3’ MSCV modules to be used as a lentiviral transfer plasmid after assembly. |
Our favourite composite part: pJUMP lentiviral transfer plasmid
By incorporating the required MSCV modules on the secondary restriction sites in the 3’ and 5’, we have been able to transform our basic backbone for the assembly of plasmids (pJUMP29-1A KanR TypeIIS Level 1 vector) into a transfer plasmid that can be used right away after the assembly. Our favorite composite part, BBa_K4501022, is suitable for lentiviral vectors of both 2nd and 3rd generation. With that, we have achieved a backbone that is suitable for both assembly and as transfer plasmid, which shortens the time and lowers the number of procedures required to assemble de constructions of interest on lentiviral vectors.
