Question (2039)
Question
I bought abm’s CRISPR gene knock out vectors/virus. Following transfection or transduction I performed sequencing and determined no gene editing had occurred. Why?
ABM community
Verified customer
Asked on Mar 24 2025
Answer
There could be several possible reasons for this:
1.. Cells are a mixture of clones, not a single clone. We recommend isolating single cells by serial dilution and then screening and sequencing those cells.
2. CRISPR gene knock out efficiency is variable and dependent on many different factors (sgRNA, cell type, chromatin accessibility, expression levels of Cas9 etc) therefore it is recommended to screen 50-100 clones in order to identify positive edited clones.
3. We also recommend abm’s CRISPR Screening Kits (Cat.No. G990 and G932) to increase your accuracy and minimize false positives and negatives.
1.. Cells are a mixture of clones, not a single clone. We recommend isolating single cells by serial dilution and then screening and sequencing those cells.
2. CRISPR gene knock out efficiency is variable and dependent on many different factors (sgRNA, cell type, chromatin accessibility, expression levels of Cas9 etc) therefore it is recommended to screen 50-100 clones in order to identify positive edited clones.
3. We also recommend abm’s CRISPR Screening Kits (Cat.No. G990 and G932) to increase your accuracy and minimize false positives and negatives.
ABM Scientific Support
Answered on Mar 24 2025