Part:BBa_K3385008

crRNA_glaA_KO_up

About the part: crRNA for upstream targeting of the glucoamylase gene (glaA) in A. niger.

Functionality: This part works with the CRISPR pFC330 [1] vector to mediate double stranded break.

Results: Below is a picture showing a tissue PCR of strains transformed with primers specific for upstream and downstream regions. Successful deletion is observed as a band at 100 bp. The ATCC 1015 strain is used as positive control and yields a band at around 2500 bp. As a negative control MQ water is used.

Tissue PCR of strains deleted with CRISPR construct.

Sequence and Features

Assembly Compatibility:

• 10
  COMPATIBLE WITH RFC[10]
• 12
  COMPATIBLE WITH RFC[12]
• 21
  COMPATIBLE WITH RFC[21]
• 23
  COMPATIBLE WITH RFC[23]
• 25
  COMPATIBLE WITH RFC[25]
• 1000
  COMPATIBLE WITH RFC[1000]

References:
[1] A CRISPR-Cas9 System for Genetic Engineering of Filamentous Fungi. Nodvig CS, Nielsen JB, Kogle ME, Mortensen UH. PLoS One. 2015 Jul 15;10(7):e0133085. doi: 10.1371/journal.pone.0133085. eCollection 2015. PONE-D-15-11561 [pii] PubMed 26177455