30. After targeting a region (named 22-24) for an internal deletion (deletion ta

Question

30. After targeting a region (named 22-24) for an internal deletion (deletion target "23") by CRISPR/Cas9, a PCR was performed on cells from the culture with guide RNAs targeting 23 ("+" in figure below, three different trials) and with no guide RNA ("-" in figure below) with the primers designed to sit on either side of the deletion target. The amplified product was run on a gel and the amplicon sizes analyzed. 22-24 423 Was the CRISPR/Cas9 successful in all the cells? Why is the band labeled d23 smaller than the band labeled 22-24?

Explanation / Answer

1) CRSIPR process targets a specific gene an with help of CAS9 which is of helicase activity cleaves specific part of DNA and inserts host DNA or other DNA sequences the experiment after excision of the target gene 23 the fragment DNA is seen in the last row, all the experiment show that the target gene is cleaved properly as after excision of target gene 23 it became a shorter sequence and lighter in weight hence it traveled longer distance than the 22-24 sequence in the row 2 which is of heavier in weight and travelled shorter distance.

2) The band 23 is of smaller weight after excision from the larger DNA molecule and forming a shorter band by travelling a longer distance while the labeled band22-24 is of larger portion and with heavy weight it travelled a shorter distance on the agarose gel electrophoresis.

Related Questions

Navigate

• Browse (All)
• Browse 3
• Subjects

---

---

Integrity-first tutoring: explanations and feedback only — we do not complete graded work. Learn more.