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Table 2 Efficiencies of correct replacements of target genes with the UDH expression cassette in multiplexed CRISPR/Cas9 using RNP complexes

From: Development of microtiter plate scale CRISPR/Cas9 transformation method for Aspergillus niger based on in vitro assembled ribonucleoprotein complexes

Target genes

gRNA

Colonies screened

Correct ΔgaaA

Correct Δ39114

Correct ΔgaaX

All correct

Targeting efficiency (%)

gaaA

gERA-009

2

2

–

–

2

100

gERA-010

5

5

–

–

5

100

gERA-009 + gERA-010

5

5

–

–

5

100

gaaA + 39114

gERA-009 + gERA-015

4

3

1

–

1

25

gERA-010 + gERA-016

7

1

2

–

0

0

gERA-009 + gERA-015+

8

1

3

–

1

13

gERA-010 + gERA-016

      

gaaA + 39114 + gaaX

gERA-001 + gERA-009 + gERA-015

6

3

1

2

0

0

gERA-002 + gERA-010 + gERA-016

6

1

1

4

1

17

gERA-001 + gERA-009 + gERA-015 + gERA-002 + gERA-010 + gERA-016

5

2

2

2

0

0

  1. Two different crRNAs for each of the target genes were designed and tested in different combinations