Background: The most prevalent type of inherited intellectual disability worldwide is Down syndrome. Prenatal testing can determine the possibility of a pregnant woman giving birth to an infant with Down’s. Many invasive interventions help with early diagnosis of Down’s. Although high-risk pregnancies should be offered invasive prenatal diagnosis, the International Society for Prenatal Diagnosis encourages non-invasive prenatal screening testing (NIPT) as a primary screening test for all pregnant women, regardless of risk. The present study aimed to determine the knowledge and attitude regarding NIPT among Saudi Arabian women. Methods: This cross-sectional study involving 1028 participants living in Saudi Arabia was conducted from February to March 2023, using a re-designed questionnaire shared among women with pregnancy history. The questionnaire assessed their knowledge, attitudes, and beliefs regarding NIPT. Results: Only 14.8% of our respondents had undergone NIPT; the most common indication was a recommendation from their healthcare provider (41.7%). About 3.4% of the respondents had given birth to a child with Down syndrome, and 22% had relatives with chromosomal abnormalities or genetic diseases. Surprisingly, only 22.3% of the respondents had previously heard about NIPT. The mean knowledge score about NIPT was 2.82 \pm 1.89. The mean knowledge score was found to be significantly higher among females who had undergone NIPT, were aged 30, and had high school education. Conclusions: Most Saudi Arabian women have poor knowledge about NIPT and a positive attitude toward it. Thus, the suggestion is to raise awareness about NIPT use, indications, prices, and possible complications.