トップページ >
CRISPR toolbox
お問い合わせ・連絡先
東邦大学理学部
生物学科
分子発生生物学研究室
村本哲哉
〒274-8510
千葉県船橋市三山2-2-1
Tel： 047-472-5165
CRISPR toolbox in Dictyostelium
Browse CRISPR vectors by application
All of the plasmids and predicted sequences are available from the National BioResource Project "Cellular Slime Molds" (NBRP Nenkin).
Knockout (SpCas9)
| Plasmid | Description | Other Information |
| SpCas9-based double-strand breaks | ||
| pTM1285 |
All-in-one CRISPR/Cas9 vector
Cas9-NLS-GFP, tRNA-sgRNA (BpiI)
Transient expression (Neo)
|
Ideal for gene knockout |
| pTM1599 | All-in-one CRISPR/Cas9 vector
Cas9-NLS, tRNA-sgRNA (BpiI)
Transient expression (Neo)
|
Ideal for gene knockout |
| pTM1756 | All-in-one CRISPR/Cas9 vector
Cas9-NLS, tRNA-sgRNA (Esp3I)
Transient expression (Hyg)
|
Ideal for knockout in G418-resistant cells |
Knockout (SpRY and SpCas9-NG)
| Plasmid | Description | Other Information |
| SpRY and SpCas9-NG based double-strand breaks | ||
| pTM1668 |
All-in-one CRISPR/Cas9 vector
SpRY-NLS, tRNA-sgRNA (Esp3I)
Transient expression (Neo)
|
Ideal for gene knockout
Genome editing at NR PAM sequences (R= A and G)
|
| pTM1718 |
All-in-one CRISPR/Cas9 vector
SpCas9-NG-NLS, tRNA-sgRNA (BpiI)
Transient expression (Neo)
|
Ideal for gene knockout
Genome editing at NG PAM sequences
|
| pTM1825 |
All-in-one CRISPR/Cas9 vector
SpRY-NLS, tRNA-sgRNA (Esp3I)
Transient expression (Hyg)
|
Ideal for knockout in G418-resistant cells
Genome editing at NR PAM sequences (R= A and G)
|
Knockout (Cas9 nickase)
| Plasmid | Description | Other Information |
| Cas9 nickase based single-strand breaks | ||
| pTM1544 | All-in-one CRISPR/Cas9 vector
Cas9 nickase-NLS-GFP, dual-tRNA-sgRNA (BpiI)
Transient expression (Neo)
|
Ideal for large deletion or knockouts
Lower off-target
Single-step cloning of dual sgRNAs
|
| pTM1866 |
All-in-one CRISPR/Cas9 vector
Cas9 nickase-NLS-GFP, dual-tRNA-sgRNA (Esp3I)
Transient expression (Hyg)
|
Ideal for gene knockout and large deletion
Lower off-target
Single-step cloning of dual sgRNAs
|
Double-knockout
| Plasmid | Description | Other Information |
| Double-knockout via one-step cloning | ||
| pTM1748 |
All-in-one CRISPR/Cas9 vector
SpRY-NLS, dual-tRNA-sgRNA (Esp3I)
Transient expression (Neo)
|
Ideal for double knockout
Genome editing at NR PAM sequences (R= A and G)
Single-step cloning of dual sgRNAs
|
| pTM1859 |
All-in-one CRISPR/Cas9 vector
Cas9-NLS, dual-tRNA-sgRNA (Esp3I)
Transient expression (Neo)
|
Ideal for double knockout
Single-step cloning of dual sgRNAs
|
Multiple gene knockout using multiplex sgRNA vector
| Plasmid | Description | Other Information |
| Multiple gene knockout | ||
| pTM1250 |
All-in-one CRISPR/Cas9 vector with pik1-5 target
Transient expression (Neo)
|
pik1-5 targets in pTM1285
multiplex CRISPR/Cas9 knockout system
|
| pTM1290 | Multiplex CRISPR/Cas9 knockout system
CRISPR final destination vector
|
Ideal for multiple gene knockouts |
| pTM1286-1289 | Multiplex CRISPR/Cas9 knockout system
tRNA-sgRNA entry vector
|
Ideal for multiple gene knockouts |
| pTM1168-1178 |
Multiplex CRISPR/Cas9 knockout system
Second module vecto
|
Ideal for multiple gene knockouts |
Inducible knockout
| Plasmid | Description | Other Information |
| Conditional knockout | ||
| pTM1670 | All-in-one CRISPR/Cas9 vector
Tet inducible Cas9-NLS, tRNA-sgRNA (Esp3I)
Transient expression (Neo)
|
Ideal for conditional gene knockout
Doxycycline-controlled inducible Cas9 system
|
| pTM1676 |
All-in-one CRISPR/Cas9 vector
Tet inducible Cas9-NLS, tRNA-sgRNA (Esp3I)
Transient expression (Blast)
|
Ideal for conditional gene knockout
Doxycycline-controlled inducible Cas9 system
|
Knock-down
| Plasmid | Description | Other Information |
| dCas9-based knock-down | ||
| pTM1765 |
All-in-one CRISPR/Cas9 vector
dCas9-NLS, dual-tRNA-sgRNA (Esp3I)
Stable expression (G418)
|
Lack of endonuclease activity
Inhibition of gene expression
Single-step cloning of dual sgRNAs
|
| pTM1870 | All-in-one CRISPR/Cas9 vector
dCas9-NLS, dual-tRNA-sgRNA (Esp3I)
Stable expression (Hyg)
|
Lack of endonuclease activity
Inhibition of gene expression
Single-step cloning of dual sgRNAs
|
Knock-in
| Plasmid | Description | Other Information |
| Knock-in with donor DNA | ||
| pTM1599 | All-in-one CRISPR/Cas9 vector
Cas9-NLS, tRNA-sgRNA (BpiI)
Transient expression (Neo)
|
Ideal for gene knockout |
| pTM1668 |
All-in-one CRISPR/Cas9 vector
SpRY-NLS, tRNA-sgRNA (Esp3I)
Transient expression (Neo)
|
Ideal for gene knockout
Genome editing at NR PAM sequences (R= A and G)
|
| pTM1718 |
All-in-one CRISPR/Cas9 vector
SpCas9-NG-NLS, tRNA-sgRNA (BpiI)
Transient expression (Neo)
|
Ideal for gene knockout
Genome editing at NG PAM sequences
|
Point-mutation
| Plasmid | Description | Other Information |
| Point-mutation with ssODN | ||
| pTM1544 | All-in-one CRISPR/Cas9 vector
Cas9 nickase-NLS-GFP, dual-tRNA-sgRNA (BpiI)
Transient expression (Neo)
|
Ideal for large deletion or knockouts
Lower off-target
Single-step cloning of dual sgRNAs
|
| pTM1748 |
All-in-one CRISPR/Cas9 vector
SpRY-NLS, dual-tRNA-sgRNA (Esp3I)
Transient expression (Neo)
|
Ideal for double knockout
Genome editing at NR PAM sequences (R= A and G)
Single-step cloning of dual sgRNAs
|
| pTM1859 |
All-in-one CRISPR/Cas9 vector
Cas9-NLS, dual-tRNA-sgRNA (Esp3I)
Transient expression (Neo)
|
Ideal for double knockout
Single-step cloning of dual sgRNAs
|