CRISPR-Cas9 Targeting

⚙️ Parameters

Guide Position 0

Mismatch Tolerance (%) 10

Specificity Weight 0.7

📊 Live Stats

—

PAM Sites

—

Guide Score

—

Off-Target Risk

—

Cleavage Efficiency

20 nt

Guide Length

NGG

PAM Motif

—

GC Content

—

On-Target Score

Target DNA Sequence

🎯 Guide RNA

Sequence—

GC Content—

Secondary Score—

Overall Score—

⚠️ Off-Target Analysis

Off-target sites—

High risk (>80%)—

Medium risk (50-80%)—

Low risk (<50%)—

Real-world connection: Cas9 recognizes NGG PAM sites (SpCas9). Guide design balances on-target activity (GC 40-60%, no poly-T) with minimal off-target hits. Off-target effects are the primary safety concern in therapeutic CRISPR applications.