Question #3: CRISPR has been used to cure an individual from sickle cell. Below is a Sanger electropherogram of a sequence from a patient without sickle cell and one with sickle cell. Sequence from a normal individual M GIC A CCT GAC T C C TGAGGAGI an AT GIG Sequence from the diseased individual n ATGGT G||C A CCT GAC TC C TG a) Where is the change in the sequence and what is the consequence to the protein sequence of this mutation? b) Below is an image of the normal and diseased quaternary hemoglobin protein. What is different about the protein shape and why does that structure have a huge impact on its function (please name the function!)? Adult haemoglobi Structure a B SCIENCEPH A GGAG SCIENCE SCIENCEphoto Sickle Cell haemoglobin S Structure a ps COLIBRARY ARY c) If you were to use CRISPR to modify the genome of a diseased individual, to which nucleotides might you design your guide RNA? Why?

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Only need help with parts a,b and c please, and thank you!

Question #3: CRISPR has been used to cure an individual from sickle cell. Below is a Sanger
electropherogram of a sequence from a patient without sickle cell and one with sickle cell.
Sequence from a normal individual
bask
AT GIGT G||C A CCT GAC с
ATGGT G C A CCT GAC TC
chr487529947
chr9:109570965
chr7:132754767
chr2203801137
a) Where is the change in the sequence and what is the consequence to the
protein sequence of this mutation?
b)
Below is an image of the normal and diseased quaternary hemoglobin
protein. What is different about the protein shape and why does that
structure have a huge impact on its function (please name the function!)?
On-target
chr465163142 TC
chr1:199581904 TAT
chr2247094773.
chrX:155356680
chrX155484568
chr2245689924.
ATACAAAGGATCA GAAT CC CNG G
CCN G
G
chr3:107039143
chr9:122015027
chr7:147113997
chrX:12851965
chrX:102677950.
Sequence from the diseased individual
sh
TG
chr5:60703887
chr5:12506322
chr4:116565656
chr10:10467604 T C
chrX:1442688686.
chr7:146700912 CA.
chr230757626
chr2:13854278
chr1573766400
C
с
chrX:13727578 .C.
chr7:143114275 C.
chr2:14708587 .C C.
chr823569322 . сст
G
.
Adult haemogloRAR
A
Structure aß,
22
c) If you were to use CRISPR to modify the genome of a diseased individual,
to which nucleotides might you design your guide RNA? Why?
d) RNA Seq is used to determine off-target effects of Cas9 cleavage. Why is this
an appropriate tool to determine these effects?
chr2216830092. сс
chr20:17814888 T
chr8:135042874 C G
SCIENCEph
e) Data on the following page is RNA-Seq data about indels that were detected
away from the target site. What does this show about the effectiveness of
CRISPR? Please also comment about the impact of these off-targets
impacts on the utility of CRISPR in human medicine.
G C
GT
GG. C-1
TT
с
- CAT
A
C.T. C
с
с
с
.
Reads
1130
621
603
406
325
320
311
149
139
139
105
103
103
85
60
53
51
49
m
TGAGGAGI
49
47
45
44
43
SCIENCE
SCIENCEphoto
42
36
35
30
M
GGA G
27
25
18
14
13
Sickle Cell haemoglobin S
Structure a s
COLIBRARY
ANVE
Transcribed Image Text:Question #3: CRISPR has been used to cure an individual from sickle cell. Below is a Sanger electropherogram of a sequence from a patient without sickle cell and one with sickle cell. Sequence from a normal individual bask AT GIGT G||C A CCT GAC с ATGGT G C A CCT GAC TC chr487529947 chr9:109570965 chr7:132754767 chr2203801137 a) Where is the change in the sequence and what is the consequence to the protein sequence of this mutation? b) Below is an image of the normal and diseased quaternary hemoglobin protein. What is different about the protein shape and why does that structure have a huge impact on its function (please name the function!)? On-target chr465163142 TC chr1:199581904 TAT chr2247094773. chrX:155356680 chrX155484568 chr2245689924. ATACAAAGGATCA GAAT CC CNG G CCN G G chr3:107039143 chr9:122015027 chr7:147113997 chrX:12851965 chrX:102677950. Sequence from the diseased individual sh TG chr5:60703887 chr5:12506322 chr4:116565656 chr10:10467604 T C chrX:1442688686. chr7:146700912 CA. chr230757626 chr2:13854278 chr1573766400 C с chrX:13727578 .C. chr7:143114275 C. chr2:14708587 .C C. chr823569322 . сст G . Adult haemogloRAR A Structure aß, 22 c) If you were to use CRISPR to modify the genome of a diseased individual, to which nucleotides might you design your guide RNA? Why? d) RNA Seq is used to determine off-target effects of Cas9 cleavage. Why is this an appropriate tool to determine these effects? chr2216830092. сс chr20:17814888 T chr8:135042874 C G SCIENCEph e) Data on the following page is RNA-Seq data about indels that were detected away from the target site. What does this show about the effectiveness of CRISPR? Please also comment about the impact of these off-targets impacts on the utility of CRISPR in human medicine. G C GT GG. C-1 TT с - CAT A C.T. C с с с . Reads 1130 621 603 406 325 320 311 149 139 139 105 103 103 85 60 53 51 49 m TGAGGAGI 49 47 45 44 43 SCIENCE SCIENCEphoto 42 36 35 30 M GGA G 27 25 18 14 13 Sickle Cell haemoglobin S Structure a s COLIBRARY ANVE
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